JP7117293B2 - TRAF6 inhibitor - Google Patents

Description

The TRAF protein family includes seven members, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5, TRAF6 and TRAF7, many of which show homology in their domain structures (Xie, P., TRAF molecules in cell signaling and in human diseases. J Mol Signal, 2013. 8(1): p. 7 and Zotti, T., P. Vito, and R. Stilo, The seventh ring: exploring TRAF7 functions. J Cell Physiol, 2012. 227 (3): p. 1280-4). Research over the last few years has elucidated a large family of signaling receptors, including TRAF proteins for signal progression. These pathways include Toll-like receptors (TCRs), IL-1 receptor family, T-cell receptors, IL-17 receptors, TNF receptors and others. TRAF-dependent signaling pathways primarily promote activation of (i) the transcription factor NF-κB and (ii) mitogen-activated protein kinase (MAPK), thereby promoting survival, proliferation, differentiation and activation. , contributes to the regulation of various cellular processes, such as cytokine production and autophagy (Xie, P., TRAF molecules in cell signaling and in human diseases. J Mol Signal, 2013. 8(1): p. 7) . Thus, distinct alterations in TRAF family members have been shown to contribute to the pathogenesis of individual human diseases, including autoimmunity, immunodeficiency and cancer (Hildebrand, J.M., Z. Yi, C.M. Buchta, et al. ., Roles of tumor necrosis factor receptor associated factor 3 (TRAF3) and TRAF5 in immune cell functions. Immunol Rev, 2011. 244(1): p. 55-74; Namjou, B., C.B. Choi, I.T. Harley, et al ., Evaluation of TRAF6 in a large multiancestral lupus cohort. Arthritis Rheum, 2012. 64(6): p. 1960-9 and Netea, M.G., C. Wijmenga, and L.A. O'Neill, Genetic variation in Toll-like receptors and disease susceptibility. Nat Immunol, 2012. 13(6): p. 535-42).

The TRAF6 protein functions both as an adapter protein and an E3 ubiquitin ligase and activates the transcription factor NF-κB. In particular, TRAF6 exhibits a RING domain and four zinc finger domains at its N-terminus, while the C-terminal TRAF domain contains a coiled-coil and a conserved MATH domain (Yin, Q., S.C. Lin, B. Lamothe, et al., E2 interaction and dimerization in the crystal structure of TRAF6. Nat Struct Mol Biol, 2009. 16(6): p. 658-66). TRAF6 directly interacts with the heterodimeric E2 enzyme Ubc13/Uev1a at its N-terminus (RING and zinc finger 1) to attach K63-linked ubiquitin chains to its substrate proteins. This process is related to T-cell receptor-mediated NF-κB activation (Oeckinghaus, A., E. Wegener, V. Welteke, et al., Malt1 ubiquitination triggers NF-kappaB signaling upon T-cell activation. EMBO J, 2007. 26(22): p. 4634-45) or innate immune responses involving IL-1β and Toll-like receptor signaling (Bhoj, V.G. and Z.J. Chen, Ubiquitylation in innate and adaptive immunity. Nature, 2009. 458( 7237): p. 430-7) are important for the progression of signaling in several cell signaling processes. Furthermore, TRAF6 is involved in IL-17 receptor signaling as well as DNA damage response (Xie, P., TRAF molecules in cell signaling and in human diseases. J Mol Signal, 2013. 8(1): p. 7 and Walsh, M.C., J. Lee, and Y. Choi, Tumor necrosis factor receptor- associated factor 6 (TRAF6) regulation of development, function, and homeostasis of the immune system. Immunol Rev, 2015. 266(1): p 72-92). Overexpression of TRAF6 and enhanced TRAF6 activity have been shown to promote chronic immune stimulation and cytokine secretion, thereby causing a wide variety of disorders, including autoimmune diseases, inflammation and cancer. Taken together, targeting TRAF6 activity by interfering with TRAF6-Ubc13 binding represents a highly attractive novel strategy for reversing disease formation.

Previous studies have focused primarily on inhibiting protein-protein interactions (PPIs) between E3 ligases and their substrates to reduce ubiquitination; p53-Mdm2 being the most prominent example. (Vassilev, L.T., B.T. Vu, B. Graves, et al., In vivo activation of the p53 pathway by small-molecule antagonists of MDM2. Science, 2004. 303(5659): p. 844-8). International patent application WO2011/160016A2 relates to isolated protein fragments containing binding pockets or active sites on E3 ligases that modulate the E2-E3 interface, and agents that interact with such binding pockets. However, WO2011/160016A2 only mentions two agents called CRIN-1 and CRIN-2 for such purposes.

Thus, there is a great need for novel, first-in-class inhibitor scaffolds that can inhibit TRAF6, particularly TRAF6 E3 ligase activity, by interfering with its interaction with the E2 enzyme Ubc13. .

The inventors of the present invention have conducted intensive research and have surprisingly found that the compounds according to Formula I, Formula II, Formula III and Formula IV, described in more detail below, meet this need. . Without wishing to be bound by theory, the inventors believe that the compounds according to the invention target the TRAF6 (E3 ligase)-Ubc13 (E2 enzyme) protein-protein interaction, thereby enhancing this biological Considered to block the pathway (E2-E3 inhibitors). In particular, this compound is believed to abolish the catalytic activity of TRAF6, thereby resulting in decreased NF-κB activation.

Compounds according to the invention are effective in reducing NF-κB activation in cell lines (see FIGS. 4 and 6) and primary T-cells derived from BALBc mice (see FIG. 8). Furthermore, we found that the compounds according to the invention improved the outcome of rheumatoid arthritis disease in the CIA mouse model (see Figure 13) and the outcome of psoriasis in the IMQ-induced psoriasis mouse model (Figure 13). 12), selectively killed ABC-DLBCL with chronic MYD88 signaling (see FIG. 14), reduced weight gain in a T2D mouse model, and also reduced IL-1β expression. (See Figure 11), reduced proliferation of U2OS cells to the same extent as 2 Gy irradiation, and application of the compound along with irradiation resulted in a synergistic effect (see Figure 15), and the compound , to affect immunoreceptor signaling in primary human peripheral blood mononuclear cells (see Figure 7). Without wishing to be bound by theory, it is believed that on a molecular basis, compounds of the present application interfere with TRAF6-Ubc13 binding (see Discussion).

（式中、
Ｘは、Ｎ及びＣＨからなる群より選択され；
Ｒ^１は、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル、（Ｃ_３－Ｃ_８）シクロアルキル、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール及び（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
Ｒ^２及びＲ^３は、独立して、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル及び（Ｃ_３－Ｃ_８）シクロアルキルからなる群より選択され；
ｎは、０と３の間の整数であり；
Ｚは、Ｃ＝Ｏ、Ｃ＝Ｓ及びＣＨ_２からなる群より選択され；
Ａは、－Ｎ（Ｒ^４）（Ｒ^５）及び下記式：

の基からなる群より選択され；
Ｒ^４は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^５は、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）ヘテロアルキル及び（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）ヘテロアルキルからなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^６は、（Ｃ_１－Ｃ_４）アルキル又は（Ｃ_１－Ｃ_４）ハロアルキルであり；
Ｙは、Ｎ－Ｂ、ＣＨ－Ｂ及びＯからなる群より選択され；
Ｂは、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）アルキル及びＮ（Ｒ^７）（Ｒ^８）からなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^７は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^８は、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
ｐは、１と２の間の整数であり；
ｑは、１と３の間の整数である）
による構造を有する化合物、又はその薬学的に許容される塩、溶媒和物若しくは水和物を提供する。 Accordingly, the present invention provides, in a first aspect, a compound for use in the treatment of cancer, immune disorders, Parkinson's disease, cardiac hypertrophy or type 2 diabetes, comprising formula I:

(In the formula,
X is selected from the group consisting of N and CH;
R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) selected from the group consisting of aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl;
R ² and R ³ are independently selected from the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl;
n is an integer between 0 and 3;
Z is selected from the group consisting of C=O, C=S and _CH2 ;
A is —N(R ⁴ )(R ⁵ ) and the following formula:

selected from the group consisting of
R ⁴ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ ;
R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl;
Y is selected from the group consisting of NB, CH-B and O;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl and N(R ⁷ )(R ⁸ ) which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ ;
R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl ( selected from the group consisting of C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl;
p is an integer between 1 and 2;
q is an integer between 1 and 3)
or a pharmaceutically acceptable salt, solvate or hydrate thereof.

（式中、
Ｘは、Ｎ及びＣＨからなる群より選択され；
Ｒ^１は、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル、（Ｃ_３－Ｃ_８）シクロアルキル、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール及び（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
Ｒ^２及びＲは、独立して、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル及び（Ｃ_３－Ｃ_８）シクロアルキルからなる群より選択され；
ｎは、０と３の間の整数であり；
Ｚは、Ｃ＝Ｏ、Ｃ＝Ｓ及びＣＨ_２からなる群より選択され；
Ａは、－Ｎ（Ｒ^４）（Ｒ^５）及び下記式：

の基からなる群より選択され；
Ｒ^４は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^５は、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）ヘテロアルキル及び（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）ヘテロアルキルからなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^６は、（Ｃ_１－Ｃ_４）アルキル又は（Ｃ_１－Ｃ_４）ハロアルキルであり；
Ｙは、Ｎ－Ｂ、ＣＨ－Ｂ及びＯからなる群より選択され；
Ｂは、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）アルキル及びＮ（Ｒ^７）（Ｒ^８）からなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^７は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^８は、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
ｐは、１と２の間の整数であり；
ｑは、１と３の間の整数である）
による構造を有する化合物、又はその薬学的に許容される塩、溶媒和物若しくは水和物を含む、医薬組成物を提供する。 In a second aspect, the present invention provides a pharmaceutical composition for use in the treatment of cancer, immune disorders, Parkinson's disease, cardiac hypertrophy or type 2 diabetes, comprising formula I:

(In the formula,
X is selected from the group consisting of N and CH;
R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) selected from the group consisting of aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl;
R ² and R are independently selected from the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl;
n is an integer between 0 and 3;
Z is selected from the group consisting of C=O, C=S and _CH2 ;
A is —N(R ⁴ )(R ⁵ ) and the following formula:

selected from the group consisting of
R ⁴ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ ;
R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl;
Y is selected from the group consisting of NB, CH-B and O;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl and N(R ⁷ )(R ⁸ ) which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ ;
R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl ( selected from the group consisting of C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl;
p is an integer between 1 and 2;
q is an integer between 1 and 3)
or a pharmaceutically acceptable salt, solvate or hydrate thereof.

（式中、
Ｘは、Ｎ及びＣＨからなる群より選択され；
Ｒ^１は、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル、（Ｃ_３－Ｃ_８）シクロアルキル、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール及び（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
Ｒ^２及びＲ^３は、独立して、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル及び（Ｃ_３－Ｃ_８）シクロアルキルからなる群より選択され；
ｎは、０と３の間の整数であり；
Ｚは、Ｃ＝Ｏ、Ｃ＝Ｓ及びＣＨ_２からなる群より選択され；
Ａは、－Ｎ（Ｒ^４）（Ｒ^５）及び下記式：

の基からなる群より選択され；
Ｒ^４は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^５は、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）ヘテロアルキル及び（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）ヘテロアルキルからなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換されており；
Ｒ^６は、（Ｃ_１－Ｃ_４）アルキル又は（Ｃ_１－Ｃ_４）ハロアルキルであり；
Ｙは、Ｎ－Ｂ、ＣＨ－Ｂ及びＯからなる群より選択され；
Ｂは、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）アルキル及びＮ（Ｒ^７）（Ｒ^８）からなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^７は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^８は、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
ｐは、１と２の間の整数であり；
ｑは、１と３の間の整数である）
による構造を有する化合物、又はその薬学的に許容される塩、溶媒和物若しくは水和物を提供する。 In a third aspect, the present invention provides a compound for use in medicine, having formula I:

(In the formula,
X is selected from the group consisting of N and CH;
R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) selected from the group consisting of aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl;
R ² and R ³ are independently selected from the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl;
n is an integer between 0 and 3;
Z is selected from the group consisting of C=O, C=S and _CH2 ;
A is —N(R ⁴ )(R ⁵ ) and the following formula:

selected from the group consisting of
R ⁴ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ ;
R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl;
Y is selected from the group consisting of NB, CH-B and O;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl and N(R ⁷ )(R ⁸ ) which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ ;
R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl ( selected from the group consisting of C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl;
p is an integer between 1 and 2;
q is an integer between 1 and 3)
or a pharmaceutically acceptable salt, solvate or hydrate thereof.

（式中、
Ｘは、Ｎ及びＣＨからなる群より選択され；
Ｒ^１は、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル、（Ｃ_３－Ｃ_８）シクロアルキル、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール及び（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、好ましくは水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_８）シクロアルキル、（Ｃ_６－Ｃ_１０）アリール及び（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
Ｒ^２及びＲ^３は、独立して、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル及び（Ｃ_３－Ｃ_８）シクロアルキル、好ましくは（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）ハロアルキルからなる群より選択され；
ｎは、０と３の間の整数であり、好ましくは１～３であり、より好ましくは１～２であり、さらにより好ましくは２であり；
Ｚは、Ｃ＝Ｏ、Ｃ＝Ｓ及びＣＨ_２からなる群より選択され、好ましくはＣ＝Ｏであり；
Ｙは、Ｎ又はＣＨであり；
Ｂは、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）アルキル及びＮ（Ｒ^７）（Ｒ^８）からなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６、好ましくはハロゲン及びＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^７は、水素又は（Ｃ_１－Ｃ_６）アルキル、好ましくは水素であり；
Ｒ^８は、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）アルキル、好ましくは（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルからなる群より選択され、より好ましくは（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキルであり；
ｐは、１と２の間の整数であり；
ｑは、１と３の間の整数である）
による構造を有する化合物、又はその薬学的に許容される塩、溶媒和物若しくは水和物である。 In some embodiments, the compound having a structure according to Formula I is represented by Formula (II):

(In the formula,
X is selected from the group consisting of N and CH;
R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably hydrogen, (C ₁ -C ₆ )alkyl , (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl;
R ² and R ³ are independently hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl, preferably (C ₁ -C ₆ ) selected from the group consisting of alkyl and (C ₁ -C ₆ )haloalkyl;
n is an integer between 0 and 3, preferably 1-3, more preferably 1-2, even more preferably 2;
Z is selected from the group consisting of C=O, C=S and _CH2 , preferably C=O;
Y is N or CH;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl and N(R ⁷ )(R ⁸ ) which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably substituted by one or more identical or different substituents selected from the group consisting of halogen and OR ⁶ ;
R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl, preferably hydrogen;
R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl ( C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl, preferably (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ ) selected from the group consisting of alkyl, more preferably (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl;
p is an integer between 1 and 2;
q is an integer between 1 and 3)
or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In some embodiments, the compound having a structure according to Formula (I) or Formula (II) is characterized in that X is N.

In some embodiments, the compound having a structure according to Formula (I) or Formula (II) is characterized in that Z is C=O.

In some embodiments, said compound having a structure according to Formula (I) or Formula (II) is
R ¹ is from hydrogen, (C ₁ -C ₄ )alkyl, (C ₃ -C ₆ )cycloalkyl, (C ₆ -C ₁₀ )aryl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl selected from the group of
R ² and R ³ are independently selected from the group consisting of (C ₁ -C ₄ )alkyl and (C ₁ -C ₄ )haloalkyl;
n is an integer between 1 and 3, preferably 1-2, more preferably 2;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl(C ₁ —C ₆ )heteroalkyl, which are optionally selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably halogen are substituted by one or more identical or different substituents.

（式中、
ｎは、１と３の間の整数であり、好ましくは１～２であり、より好ましくは２であり；
Ｙは、Ｎ又はＣＨであり；
Ｂは、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）アルキルからなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６、好ましくはハロゲン及びＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
ｑは、１～３の、好ましくは１～２の整数、より好ましくは２である；
による構造を有する化合物、又はその薬学的に許容される塩、溶媒和物若しくは水和物である。 In some embodiments, the compound having a structure according to formula (I) above has formula (III):

(In the formula,
n is an integer between 1 and 3, preferably 1-2, more preferably 2;
Y is N or CH;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably substituted by one or more identical or different substituents selected from the group consisting of halogen and OR ⁶ ;
q is an integer from 1 to 3, preferably from 1 to 2, more preferably 2;
or a pharmaceutically acceptable salt, solvate or hydrate thereof.

（式中、
Ｘは、Ｎ及びＣＨからなる群より選択され；
Ｒは、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル、（Ｃ_３－Ｃ_８）シクロアルキル、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール及び（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、好ましくは水素及び（Ｃ_１－Ｃ_６）アルキル、より好ましくは（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
Ｒ^２及びＲ^３が、独立して、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル及び（Ｃ_３－Ｃ_８）シクロアルキル、好ましくは（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）ハロアルキル、より好ましくは（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
ｎは、０と３の間の整数であり、好ましくは１～３であり、より好ましくは１～２であり、より好ましくは２であり；
Ｚは、Ｃ＝Ｏ、Ｃ＝Ｓ及びＣＨ_２からなる群より選択され、好ましくはＣ＝Ｏであり；
Ｒ^４は、水素又は（Ｃ_１－Ｃ_６）アルキル、好ましくは水素であり；
Ｒ^５は、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）ヘテロアルキル及び（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）ヘテロアルキルからなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６、好ましくはハロゲンからなる群より選択される、１つ以上の同一又は異なる置換基によって任意に置換され；
Ｒ^６は、（Ｃ_１－Ｃ_４）アルキル又は（Ｃ_１－Ｃ_４）ハロアルキルである）
による構造を有する化合物、又はその薬学的に許容される塩、溶媒和物若しくは水和物である。 In some embodiments, the compound having a structure according to formula (I) above has formula (IV):

(In the formula,
X is selected from the group consisting of N and CH;
R is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ )aryl (C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably hydrogen and (C ₁ -C ₆ )alkyl, more preferably selected from the group consisting of (C ₁ -C ₆ )alkyl;
R ² and R ³ are independently hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl, preferably (C ₁ -C ₆ ) selected from the group consisting of alkyl and (C ₁ -C ₆ )haloalkyl, more preferably (C ₁ -C ₆ )alkyl;
n is an integer between 0 and 3, preferably 1-3, more preferably 1-2, more preferably 2;
Z is selected from the group consisting of C=O, C=S and _CH2 , preferably C=O;
R ⁴ is hydrogen or (C ₁ -C ₆ )alkyl, preferably hydrogen;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , optionally substituted by one or more identical or different substituents selected from the group consisting of halogen;
R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl)
or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In some embodiments, the cancer is lymphoma such as diffuse large B-cell lymphoma (DLBCL) and MALT lymphoma, multiple myeloma (MM), lung cancer, lung adenocarcinoma, colon cancer, prostate cancer, breast cancer. , osteosarcoma, pancreatic cancer and esophageal squamous cell carcinoma (ESCC).

In some embodiments, said immune disease is an autoimmune disease, preferably psoriasis, rheumatoid arthritis, celiac disease, inflammatory bowel disease such as Crohn's disease and ulcerative colitis, multiple sclerosis and diabetes type 1. An autoimmune disease selected from the group consisting of

（式中、
Ｘは、Ｎ及びＣＨからなる群より選択され；
Ｒは、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル、（Ｃ_３－Ｃ_８）シクロアルキル、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール及び（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
Ｒ^２及びＲ^３は、独立して、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル及び（Ｃ_３－Ｃ_８）シクロアルキルからなる群より選択され；
ｎは、０と３の間の整数であり；
Ｚは、Ｃ＝Ｏ、Ｃ＝Ｓ及びＣＨ_２からなる群より選択され；
Ａは、下記式：

の基からなる群より選択され；
Ｂ^１は、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル又は（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルであり、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｂ^２は、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ_６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換されている（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキルであるか、あるいは、
Ｂ^２は、場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基で置換される（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルであり；
Ｂ^３は、水素、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）アルキル及びＮ（Ｒ^７）（Ｒ^８）からなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^６は、（Ｃ_１－Ｃ_４）アルキル又は（Ｃ_１－Ｃ_４）ハロアルキルであり；
Ｒ^７は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^８は、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
ｐは１であり、かつ
ｑは、１と２の間の整数であるか、又は、
ｐは２であり、かつ
ｑは３である）
による構造を有する化合物、又はその薬学的に許容される塩、溶媒和物若しくは水和物を提供する。 In a fourth aspect, the present invention provides a compound of formula I:

(In the formula,
X is selected from the group consisting of N and CH;
R is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ )aryl (C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl;
R ² and R ³ are independently selected from the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl;
n is an integer between 0 and 3;
Z is selected from the group consisting of C=O, C=S and _CH2 ;
A is the following formula:

selected from the group consisting of
B ¹ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl or (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ substituted by one or more identical or different substituents;
B ² is substituted by one or more same or different substituents selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ₆ ( C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl; or
B ² is optionally substituted with one or more same or different substituents selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl;
B ³ is hydrogen, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl and N(R ⁷ )(R ⁸ ), which are optionally selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , substituted by one or more identical or different substituents;
R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl;
R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl ( selected from the group consisting of C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl;
p is 1 and q is an integer between 1 and 2, or
p is 2 and q is 3)
or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In some embodiments, the compound is
1-(4-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperazin-1-yl)propan-1-one,
(R,S)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b] pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperazin-1-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
(R)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
(S)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-cyclopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5- dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-phenyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(1-(3-benzyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( 4-benzylpiperidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(4-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)ethan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propane-1-thione,
6-(4-(3-(4-benzylpiperidin-1-yl)propyl)-3,5-dimethyl-1H-pyrazol-1-yl)-3-methyl-[1,2,4]triazolo[4 , 3-b]pyridazine,
1-(azepan-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H- pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-morpholino propane-1-one,
N-(3-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)-N-methylpropanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(furan-2-ylmethyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-phenylpropan-2-yl)piperazin-1-yl)propan-1-one,
1-(4-(3,4-dimethoxybenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b ] pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(naphthalen-2-ylmethyl)piperazin-1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylazetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl )-1H-pyrazol-4-yl)propan-1-one,
1-(3-(benzylamino)azetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6 -yl)-1H-pyrazol-4-yl)propan-1-one,
N-benzyl-3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole-4-carboxamide,
(3-benzylpyrrolidin-1-yl)(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole- 4-yl)methanone,
1-(3-benzylpyrrolidin-1-yl)-3-(5-isopropyl-3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6- yl)-1H-pyrazol-4-yl)propan-1-one, and 1-(3-benzylpyrrolidin-1-yl)-3-(3-methyl-1-(3-methyl-[1,2, 4]triazolo[4,3-b]pyridazin-6-yl)-5-(trifluoromethyl)-1H-pyrazol-4-yl)propan-1-one.

In some embodiments, the compound is
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperazin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorophenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(4 -fluorophenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( pyrrolidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(3 - methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(4 - methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-phenylpiperazine -1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(pyridine -2-yl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( (5-methylfuran-2-yl)methyl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-fluorophenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 3-phenylpropyl)propanamide,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-benzylpiperidine -1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2,5-dimethylphenyl)piperazin-1-yl)propan-1-one,
N-(4-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( thiophen-2-ylmethyl)propanamide,
N-(2-(1H-indol-3-yl)ethyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propanamide,
N-benzyl-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) propanamide,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-N-(3-phenylpropyl ) propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-ethylpiperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-methylpiperidine -1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(pyrrolidin-1- yl) propane-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 2-fluorobenzyl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 2-((furan-2-ylmethyl)thio)ethyl)propanamide,
1-(4-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperazin-1-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperazin-1-yl)propan-1-one,
(R)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
(S)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-cyclopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5- dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-phenyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(1-(3-benzyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( 4-benzylpiperidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(4-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)ethan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propane-1-thione,
6-(4-(3-(4-benzylpiperidin-1-yl)propyl)-3,5-dimethyl-1H-pyrazol-1-yl)-3-methyl-[1,2,4]triazolo[4 , 3-b]pyridazine,
1-(azepan-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H- pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-morpholino propane-1-one,
N-(3-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)-N-methylpropanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(furan-2-ylmethyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-phenylpropan-2-yl)piperazin-1-yl)propan-1-one,
1-(4-(3,4-dimethoxybenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b ] pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(naphthalen-2-ylmethyl)piperazin-1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylazetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl )-1H-pyrazol-4-yl)propan-1-one,
1-(3-(benzylamino)azetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6 -yl)-1H-pyrazol-4-yl)propan-1-one,
N-benzyl-3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole-4-carboxamide,
(3-benzylpyrrolidin-1-yl)(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole- 4-yl)methanone,
1-(3-benzylpyrrolidin-1-yl)-3-(5-isopropyl-3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6- yl)-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-5 -(trifluoromethyl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxyphenyl)piperazin-1-yl)propan-1-one,
1-benzyl-4-(3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole-4 -yl)propanoyl)piperazin-2-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-diethyl-1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl -1H-pyrazol-4-yl)propan-1-one,
1-(2,2-dimethylpyrrolidin-1-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5 -dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(3,3-dimethylpyrrolidin-1-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5 -dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-diethyl-1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-diethyl-1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 3,3-dimethylpyrrolidin-1-yl)propan-1-one,
1-((1S)-2-azabicyclo[2.2.1]heptan-2-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)propan-1-one,
(1S,5R)-3-(3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazole -4-yl)propanoyl)-2,3,4,5-tetrahydro-1,5-methanopyrido[1,2-d][1,4]diazepin-7(1H)-one,
1-((1S)-2-azabicyclo[2.2.1]heptan-2-yl)-3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-(trifluoromethyl)-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) -1-(pyrrolidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl -1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 3,3-dimethylpyrrolidin-1-yl)propan-1-one,
3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( pyrrolidin-1-yl)propan-1-one,
3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( pyrrolidin-1-yl)propan-1-one,
3-(3,5-diethyl-1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 3,3-dimethylpyrrolidin-1-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-diethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(4-benzyl-1,4-diazepan-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3-isopropyl-5-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6- yl)-1H-pyrazol-4-yl)propan-1-one,
3-(1-(3-benzyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( 4-benzylpiperidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-cyclopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5- dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperazin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(3-benzylpiperazin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(3-benzylpyrrolidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenethylpiperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-(4-phenethylpiperidin-1-yl ) propan-1-one.

In another aspect, the present invention relates to a pharmaceutical composition comprising a compound having a structure according to formula (I) above, or said compound selected from the group above, and at least one pharmaceutically acceptable carrier.

In another aspect, the present invention provides a kit comprising a compound having a structure according to formula (I) above, said compound selected from the group above, or said pharmaceutical composition, and at least one pharmaceutically acceptable carrier Regarding.

Other features and advantages of the invention will become apparent from the following detailed description, drawings, and claims.

Compound identification and C25 SAR studies. (A) Identification process of TRAF6-Ubc13 inhibitors. ALPHAScreen technology was used to screen 25,000 small molecules from three in-house libraries. 520 molecules inhibited TRAF6-Ub13 binding by >25%. After removing ALPHA frequent hitters and His-tag frequent hitters, 178 compounds were retested in the 5-point titration ALPHAScreen assay. The remaining 27 small molecules were rearranged and tested in ALPHAScreen assays (10 point increments) as well as EMSA experiments. C25 showed the best results in in vitro and cell-based assays. (B) C25-140 appears equally effective in the ALPHAScreen, but is more active in the (C) cell-based ALPHASurefire experiment. C25 equals compound 1 herein, C25-140 equals compound 3 herein, C25-167 equals compound 61 herein, and C25-189 equals compound 2 herein . Biochemical and cell-based characterization of C25-140. (A) shows the chemical structure of C25-140. (B) C25-140 dose-dependently reduces the interaction of endogenous Ubc13 to ectopically expressed HA-tagged TRAF6 in HEK293T-cells. (C) C25-140 directly binds to TRAF6 RZ1 protein as evidenced by distinct peak shifts or disappearance in NMR experiments. (D) The TRAF6 inhibitor C25-140 dose-dependently reduces the ability of TRAF6 and Ubc13 to form free polyubiquitin chains in an in vitro ubiquitination assay. (E) The interaction of Ubc13 and Uev1a was not affected by C25-140. C25-140 is equivalent to compound 3 herein. Effect of C25-140 on cell viability and cell cycle. (A) C25-140 had little effect on metabolic activity and cell viability after 24 h incubation at concentrations up to 50 μM, as observed in three different cell lines (MEF, HepG2 and Jurkat-cells). do not affect Toxicity is noticeable only at the highest dose of 100 μM. (BC) Cell cycle progression in Jurkat T-cells and (DE) U2OS cells is unimpaired after 24 and 72 hours of C25-140 treatment. C25-140 is equivalent to compound 3 herein. C25-140 interferes with IL-1β and TNFα signaling in MEF cells. (A) C25-140 treatment reduces IL-1β-induced autoubiquitination of TRAF6. (B) In the IKK activity assay, C25-140 treatment causes decreased phosphorylation of IκBα following IL-1β stimulation. (C) mRNA expression of IL-1β-inducible target genes (eg, ICAM-1 and A20) is reduced by C25-140 treatment. (D) C25-140 also affects TNFα-induced phosphorylation of IκBα in the IKK activity assay and (E) mRNA expression levels of target genes ICAM-1 and A20. IKK KA means IKK kinase activity. C25-140 is equivalent to compound 3 herein. C25-140 interferes with IL-1β and TNFα signaling in MEF cells. (A) C25-140 treatment reduces IL-1β-induced autoubiquitination of TRAF6. (B) In the IKK activity assay, C25-140 treatment causes decreased phosphorylation of IκBα following IL-1β stimulation. (C) mRNA expression of IL-1β-inducible target genes (eg, ICAM-1 and A20) is reduced by C25-140 treatment. (D) C25-140 also affects TNFα-induced phosphorylation of IκBα in the IKK activity assay and (E) mRNA expression levels of target genes ICAM-1 and A20. IKK KA means IKK kinase activity. C25-140 is equivalent to compound 3 herein. (A) Using an in vitro ubiquitination assay, it can be demonstrated that self-ubiquitination of cIAP1 (RING E3 ligase; K63 chain formation) is reduced with C25-140 treatment in a dose-dependent manner. In contrast, (B) p53 ubiquitination by MDM2 and S5A ubiquitination by MURF/TRIM63 (both RING E3 ligase; K48 chain formation), (C) RNF4 autoubiquitination (SUMO targeted ubiquitin ligase), and (D) ITCH autoubiquitination and S5A ubiquitination by E6AP (both HECT E3 ligase; K48 chain formation) are unaffected by C25-140. (E) A ubiquitin profiler panel was performed. Various E2 and E3 ligases in combination with substrates were incubated with 30 μM compound C25-140. Neither reaction is significantly inhibited by compound C25-140. C25-140 is equivalent to compound 3 herein. C25-140 reduces T-cell receptor signaling in Jurkat T-cells. (A) TRAF6 autoubiquitination induced by T-cell receptor (TCR) stimulation with PMA/ionomycin (P/I) is dose-dependently reduced by C25-140 treatment. (B) IKK kinase activity is decreased after C25-140 treatment, which in turn leads to (C) reduced IκBα degradation and attenuated NF-κB activation. (DE) mRNA levels and cytokine secretion of TCR target genes IL-2 and TNFα are decreased in a dose-dependent manner. IKK KA means IKK kinase activity. C25-140 is equivalent to compound 3 herein. C25-140 affects immunoreceptor signaling in primary human PBMC. (A) IL-1β-stimulated human peripheral blood mononuclear cells (PBMC) secretion of inflammatory cytokines (eg, IL-6 and TNFα) can be attenuated by C25-140. (B) LPS-stimulated human PBMC secreted IL-1β and TNFα cytokines at reduced levels. (C) Human PBMC stimulated with CD3/CD28 to induce TCR signaling show decreased levels of IL-2 and TNFα secretion upon C25-140 treatment. C25-140 is equivalent to compound 3 herein. C25-140 interferes with TCR signaling in primary mouse T-cells. Isolated primary naive CD4 ⁺ T-cells, in the presence of C25-140, show decreased IL-2 mRNA expression and IL-2 protein secretion when stimulated with CD3/CD28. C25-140 is equivalent to compound 3 herein. ADME and pharmacokinetic studies of C25-140. (A) Absorption, distribution, metabolism and excretion (ADME) parameters of C25-140 revealed a stable and suitable small molecule for further drug development. (BC) Pharmacokinetic studies of C25-140 administered intravenously (IV), orally (PO) and intraperitoneally (IP) showed moderate in vivo half-life, rapid initial distribution phase and favorable biology. demonstrating availability. C25-140 is equivalent to compound 3 herein. SafetyScreen87 to assess the in vivo safety of compound C25-140. Here, 87 assays were performed at Eurofins Cerep Panlabs. The 87 major molecular targets include kinases, non-kinase enzymes, nuclear receptors, GPCRs, transporters and ion channels, which were tested at a compound concentration of 10 μM C25-140. C25-140 did not significantly affect any of the 87 molecular targets. Assay, ligand/substrate, species and tissue are indicated. Reaction rate is indicated. C25-140 is equivalent to compound 3 herein. C25-140 treatment of diet-induced obese (DIO) mice affects weight gain and epidermal white adipose tissue inflammation. (A) C25-140 treated mice show less weight gain compared to control mice, while (B) food intake remains unaffected in both groups. (C) mRNA levels of proinflammatory cytokines IL-1β and NF-κB dependent target gene VCAM are decreased in C25-140 treated mice. C25-140 is equivalent to compound 3 herein. Vhcl means vehicle. C25-140 ameliorates symptoms of IMQ-induced psoriasis. (A) Psoriasis mouse study design. IMQ was applied once daily and C25-140 was applied topically twice daily to the shaved back and right ear. Parameters were scored daily and samples were taken on day 6 for IL-17 cytokine measurements. (B) C25-140 decreased the cumulative score consisting of (C) dorsal thickness score, (D) desquamation score and (E) dorsal erythema score. (F) Ear thickness is significantly reduced with C25-140 treatment. (G) The abundance of cytokine IL-17 is decreased in the presence of C25-140. C25-140 is equivalent to compound 3 herein. C25-140 alleviates symptoms of collagen-induced arthritis. (A) Collagen-induced arthritis mouse study design. Rheumatoid arthritis (RA) was induced by collagen injection. On day 21, mice received a collagen boost injection to induce the development of arthritic symptoms. After 7 days, C25-140 was administered twice daily for 14 days. Mice were scored daily for arthritis index (AI) and euthanized on day 42. Limbs were collected for histopathological analysis. (B) Body weight was measured periodically during the study and showed no signs of weight loss. (C) AI in C25-140-treated mice is dose-dependently decreased compared to control mice. (DE) Limb global histological scores, including inflammation, pannus formation, cartilage damage and bone resorption, can be attenuated by C25-140 treatment. (F) Histopathology slides from the ankles and hindlimbs of each population support the results of the scored parameters. C25-140 is equivalent to compound 3 herein. C25-140 alleviates symptoms of collagen-induced arthritis. (A) Collagen-induced arthritis mouse study design. Rheumatoid arthritis (RA) was induced by collagen injection. On day 21, mice received a collagen boost injection to induce the development of arthritic symptoms. After 7 days, C25-140 was administered twice daily for 14 days. Mice were scored daily for arthritis index (AI) and euthanized on day 42. Limbs were collected for histopathological analysis. (B) Body weight was measured regularly during the study and showed no signs of weight loss. (C) AI in C25-140-treated mice is dose-dependently decreased compared to control mice. (DE) Limb global histological scores, including inflammation, pannus formation, cartilage damage and bone resorption, can be attenuated by C25-140 treatment. (F) Histopathology slides from the ankles and hindlimbs of each population support the results of the scored parameters. C25-140 is equivalent to compound 3 herein. C25-140 reduces viability of MYD88-dependent ABC-DLBCL cells. Viability of MYD88-dependent ABC-DLBCL (HBL1) is reduced by 30 μM C25-140 treatment, whereas MYD88 wild-type ABC-DLBCL (U2932) and germinal center B-cell (GCB) DLBCL (BJAB) are C25-140 Not affected by processing. Compounds were added on days 0, 4 and 8. C25-140 is equivalent to compound 3 herein. Combination therapy of irradiation and C25-140 treatment shows a synergistic effect on osteosarcoma cell proliferation. Proliferation of osteosarcoma cells is reduced by either 30 μM C25-140 treatment or 2 Gy irradiation. This combination (30 μM C25-140+2 Gy) reduces cell proliferation to the same extent as the higher irradiation dose (4 Gy). C25-140 is equivalent to compound 3 herein. Analysis of the effects of C25-140 on a panel of tumor cell lines. Seventy-eight tumor cell lines from different origins were tested for loss of viability with C25-140 treatment. Two control cell lines (HS-5 and MCF 10A) were used. Various cancer cell lines were killed by C25-140 treatment, but not control cell lines. A major effect was evident in blood-derived cancer cells, but cancer cell lines of other origins were also affected. C25-140 is equivalent to compound 3 herein. Analysis of nine structural analogues of C25-140 in the ALPHAScreen assay. Nine non-commercially available structural analogues of C25-140 were tested in serial dilution experiments using the ALPHAScreen assay. All compounds tested show inhibitory activity, with HZM06-04 showing the most pronounced effect. Importantly, none of the small molecules tested non-specifically interfered with the ALPHAScreen technology in the TruHits experiments. C25 equals compound 1, C25-140 equals compound 3, HZM06-1 equals compound 28, HZM06-2 equals compound 29, HZM06-3 equals compound 30, HZM06-4 equals compound 31 HZM06-6 equals Compound 32, HZM06-7 equals Compound 33, HZM06-8 equals Compound 34, HZM06-9 equals Compound 35, and HZM06-10 equals Compound 36 herein. equal. The Z' value is 0.77 and the signal window is 10.29. Cytotoxicity measurements of nine structural analogues of C25-140 in various cell lines. Jurkat, HepG2 and MEF cells were tested for viability when treated with the nine analogues for 6 hours or 24 hours. All small molecules show less cytotoxicity compared to C25-140. C25 equals compound 1, C25-140 equals compound 3, HZM06-1 equals compound 28, HZM06-2 equals compound 29, HZM06-3 equals compound 30, HZM06-4 equals compound 31 HZM06-6 equals Compound 32, HZM06-7 equals Compound 33, HZM06-8 equals Compound 34, HZM06-9 equals Compound 35, and HZM06-10 equals Compound 36 herein. equal. Co-immunoprecipitation studies of nine structural analogues of C25-140 reveal the inhibitory potency of individual analogues in cells. In co-immunoprecipitation studies, individual compounds including C25-140, HZM06-04, HZM06-07, HZM06-08 and HZM06-09 showed inhibition of TRAF6-Ubc13 binding in HEK293T cells. C25 equals compound 1, C25-140 equals compound 3, HZM06-1 equals compound 28, HZM06-2 equals compound 29, HZM06-3 equals compound 30, HZM06-4 equals compound 31 Equal, HZM06-6 equals compound 32, HZM06-7 equals compound 33, HZM06-8 equals compound 34, HZM06-9 equals compound 35, and HZM06-10 equals compound 36. Similar compounds. All compounds were tested in the ALPHAScreen assay. Inhibition levels are shown.

[Detailed description]
Although the present invention is described in detail below, it is understood that this invention is not limited to the particular methodology, protocols and reagents described herein, as such may vary. Also, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the invention, which is limited only by the appended claims. is also understood. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

The constituent elements of the present invention are described below. Although these elements are listed with specific embodiments, it should be understood that they can be combined in any number and in any way to create additional embodiments. The various examples and preferred embodiments described throughout the specification should not be construed to limit the invention to only the explicitly described embodiments. This description is to be understood to support and encompass embodiments that combine the explicitly described embodiments with any number of the disclosed and/or preferred elements. Moreover, any variations and combinations of all the elements described herein should be considered disclosed by the description of the present application unless the context indicates otherwise. For example, in one embodiment of compounds of the invention, when R ¹ is heteroaryl (such as pyrazolyl), and in another embodiment of compounds of the invention, B is a substituted or unsubstituted phenyl ring, a preferred practice is In embodiments, R ¹ of the compounds of the invention is heteroaryl (such as pyrazolyl) and B is a substituted or unsubstituted phenyl ring.

Throughout this specification and the claims that follow, unless the context requires otherwise, the word "comprises" and variations such as "comprises" and "comprises" refer to a stated member, integer or step, or Although it is understood to mean the inclusion of a group of members, integers or steps, and not the exclusion of any other member, integer or step, or group of members, integers or steps, in some embodiments: Such other members, integers or steps, or groups of members, integers or steps, may be excluded, i.e., the substance of the subject matter is the stated member, integer or step, or the member, integer or step. in the inclusion of the group of The terms "a" and "an" and "The" and similar references used in the context of describing the invention (particularly in the context of the claims) are defined as or should be construed to cover both the singular and the plural unless the context clearly contradicts. Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated herein as if it were individually listed herein.

All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language provided herein (eg, "such as"), is intended merely to better illustrate the invention, and may be otherwise claimed. It does not pose a limitation on the scope of the invention. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

Unless otherwise indicated, the term "at least" preceding a series of elements should be understood to refer to all elements within that series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be included in this invention.

As used herein, "consisting of" excludes any element, step, or ingredient not specified in the claim element. As used herein, "consisting essentially of" does not exclude materials or steps that do not materially affect the basic and novel features of the claim. In each case herein, any of the terms "comprising", "consisting essentially of" and "consisting of" may be replaced by either of the other two terms.

Several documents are cited throughout the text of this specification. Each document cited herein (including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc.), whether supra or infra, is hereby incorporated by reference in its entirety. incorporated. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such disclosure by virtue of prior invention.

Compounds In order that the invention may be more readily understood, some terms are first defined. Additional definitions are set forth throughout the description.

As used herein and throughout the description, the term "alkyl" refers to a saturated straight or branched chain hydrocarbon monovalent group. Preferably, the alkyl group contains 1 to 6 carbon atoms, ie 1, 2, 3, 4, 5 or 6 carbon atoms, preferably 1 to 4 carbon atoms. In another embodiment, the alkyl group employed contains 1-5 carbon atoms (C _1-5 alkyl). In another embodiment, the alkyl group employed contains 1-4 carbon atoms (C _1-4 alkyl). In another embodiment, the alkyl group employed contains 1-3 carbon atoms (C _1-3 alkyl). In another embodiment, the alkyl group employed contains 1-2 carbon atoms (C _1-2 alkyl). In another embodiment, the alkyl group employed is methyl. Examples of alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, 1,2-dimethylpropyl, isoamyl, n-hexyl, isohexyl, sec -hexyl and the like, which may have one or more substituents. Alkyl group substituents include, but are not limited to, any of the substituents described herein that result in the formation of stable moieties. Substituted alkyl groups can be substituted at any position as long as the resulting compound is sufficiently stable and suitable as a pharmaceutically active compound. The prerequisite that certain groups and compounds of any one of the formulas I, II, III and IV are sufficiently stable and suitable as pharmaceutically active compounds is generally the Applies to all groups of any one compound. If the alkyl group may be mono- or polysubstituted by fluorine, the alkyl group may be unsubstituted, i.e., may not carry fluorine atoms, or may be, for example, 1, 2, 3, 4, 5 , 6, 7, 8, 9, 10 or 11 fluorine atoms, preferably 1, 2, 3, 4 or 5 fluorine atoms, which can be present in any position. For example, in a fluoro-substituted alkyl group, one or more methyl groups can be present as trifluoromethyl groups, each having three fluorine atoms, and/or one or more methylene groups (CH ₂ ) can be present as difluoromethylene groups each having two fluorine atoms. The statements regarding substitution of a group by fluorine also apply if the group further carries other substituents and/or is part of another group, eg an alkyl-O- group. Examples of fluoro-substituted alkyl groups are trifluoromethyl, 2-fluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, 3,3,3-trifluoropropyl, 2, 2,3,3,3-pentafluoropropyl, 4,4,4-trifluorobutyl and heptafluoroisopropyl. Examples of fluoro-substituted alkyl-O- groups are trifluoromethoxy, 2,2,2-trifluoroethoxy, pentafluoroethoxy and 3,3,3-trifluoropropoxy. Examples of fluoro-substituted alkyl-S(O) _m - groups are trifluoromethanesulfanyl (CF ₃ -S-, trifluoromethylsulfanyl-), trifluoromethanesulfinyl (CF ₃ -S(O)-) and trifluoromethanesulfonyl ( CF ₃ —S(O) ₂ —). In some embodiments, the alkyl group is selected from halogen, hydroxyl, cyano, (C ₁ -C ₆ )alkyl-O- and (C ₁ -C ₆ )alkyl-S(O) _m -, 1 It may be substituted with one or more same or different substituents. Examples of (C ₁ -C ₆ )alkyl-O- are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy and n-pentoxy. Examples of alkyl-S(O) _m are methanesulfanyl (CH ₃ —S—, methylsulfanyl), methanesulfinyl (CH ₃ —S(O)—), methanesulfonyl (CH ₃ —S(O) ₂ —). , ethanesulfanyl (CH ₃ —CH ₂ —S—, ethylsulfanyl-), ethanesulfinyl (CH ₃ —CH ₂ —S(O)—), ethanesulfonyl (CH ₃ —CH ₂ —S(O) ₂ —) , 1-methylethanesulfanyl ((CH3) ₂ CH—S—, 1-methylethylsulfanyl-), 1-methylethanesulfinyl ((CH ₃ ) ₂ CH—S(O)—) and 1-methylethanesulfonyl (( CH ₃ ) ₂ CH—S(O) ₂ —). In one embodiment of the invention, the numbers m are selected from 0 and 2 and all numbers m are independent of each other and may be the same or different. For example, when an alkyl group is substituted with a CF ₃ —S— group, the CF ₃ —S— group may be substituted with sulfur, as represented by a terminal line (hyphen) next to the sulfur atom representing a free bond. is attached to this alkyl group via In some embodiments, the alkyl chain is straight. In some embodiments, the alkyl chain is branched. In some embodiments, the alkyl chain is substituted. In some embodiments, alkyl chains are unsubstituted. In some embodiments, alkyl chains are linear and substituted or unsubstituted. In some embodiments, alkyl chains are branched and substituted or unsubstituted.

As used herein and throughout the description, the term "alkylene" refers to a saturated straight or branched chain hydrocarbon divalent radical. Preferably, alkylene contains 1 to 6 carbon atoms, ie 1, 2, 3, 4, 5 or 6 carbon atoms, more preferably 1 to 4 carbon atoms. Exemplary alkylene groups are methylene, ethylene (ie, 1,1-ethylene, 1,2-ethylene), propylene (ie, 1,1-propylene, 1,2-propylene (--CH(CH ₃ )CH ₂ -), 2,2-propylene (-C(CH ₃ ) ₂ -) and 1,3-propylene), butylene isomers (e.g. 1,1-butylene, 1,2-butylene, 2,2-butylene, 1,3-butylene, 2,3-butylene (cis or trans or mixtures thereof), 1,4-butylene, 1,1-isobutylene, 1,2-isobutylene and 1,3-isobutylene), pentylene isomerism structure (for example, 1,1-pentylene, 1,2-pentylene, 1,3-pentylene, 1,4-pentylene, 1,5-pentylene, 1,1-isopentylene, 1,1-sec-pentyl, 1, 1-neopentyl), hexylene isomers (for example, 1,1-hexylene, 1,2-hexylene, 1,3-hexylene, 1,4-hexylene, 1,5)-hexylene, 1,6-hexylene and 1, 1-isohexylene), etc. Alkylene groups may be cyclic or acyclic, branched or unbranched, substituted or unsubstituted. Alkylene group substituents include, but are not limited to, any of the substituents described herein that result in the formation of stable moieties.

As used herein and throughout the description, the term "haloalkyl" refers to an alkyl group substituted with one halogen substituent up to perhalo substitution. A halogen substituent is preferably fluorine. Haloalkyl is preferably perfluoroalkyl. In some embodiments, the haloalkyl group employed in the invention contains 1-6 carbon atoms (C _1-6 haloalkyl). In another embodiment, the haloalkyl group employed contains 1-5 carbon atoms (C _1-5 haloalkyl). In another embodiment, the haloalkyl group employed contains 1-4 carbon atoms (C _1-4 haloalkyl). In another embodiment, the haloalkyl group employed contains 1-3 carbon atoms (C _1-3 haloalkyl). In another embodiment, the haloalkyl group employed contains 1-2 carbon atoms (C _1-2 haloalkyl). In another embodiment, the haloalkyl group employed contains 1 carbon atom (C ₁ haloalkyl). In another embodiment, the haloalkyl group employed is trifluoromethyl. Exemplary fluoro-substituted C ₁ -C ₂ alkyls are —CFH ₂ , —CF ₂ H, —CF ₃ , CH ₂ CH ₂ F, —CH ₂ CHF ₂ , —CHFCH ₃ , —CHFCH ₃ , —CF ₂ CHF. ₂ . Perfluoro-substituted C ₁ -C ₂ haloalkyl includes, for example, —CF ₃ and CF ₂ CF ₃ .

As used herein, throughout the description, the term "heteroalkyl" as used herein includes one or more heteroatoms (e.g., oxygen, sulfur, nitrogen, phosphorus or silicon atoms) between carbon atoms. ), as defined herein. A heteroalkyl can be substituted or unsubstituted. In certain embodiments, the heteroalkyl group contains 1-6 carbon atoms and 1-3 heteroatoms (C _1-6 heteroalkyl). In certain embodiments, the heteroalkyl group contains 1-5 carbon atoms and 1-3 heteroatoms (C _1-5 heteroalkyl). In certain embodiments, the heteroalkyl group contains 1-4 carbon atoms and 1-2 heteroatoms (C _1-4 heteroalkyl). In certain embodiments, the heteroalkyl group contains 1-3 carbon atoms and 1 heteroatom (C _1-3 heteroalkyl). In certain embodiments, the heteroalkyl group contains 1-2 carbon atoms and 1 heteroatom (C _1-2 heteroalkyl). The term "heteroalkylene," as used herein, refers to a divalent group derived by removal of two hydrogen atoms from a heteroalkyl group, as defined herein. Heteroalkylene groups may be cyclic or acyclic, branched or unbranched, substituted or unsubstituted. In certain embodiments, the heteroalkyl group is a substituted heteroalkyl group containing 1-6 carbon atoms and 1-3 heteroatoms (C _1-6 heteroalkyl). In certain embodiments, the heteroalkyl group is an unsubstituted heteroalkyl group containing 1-6 carbon atoms and 1-3 heteroatoms (C _1-6 heteroalkyl). In some embodiments, heteroalkyl is an alkyl moiety in which S is substituted for the methylene group. In some embodiments, heteroalkyl is an alkyl moiety in which the methylene group is replaced with O. In some embodiments, heteroalkyl is an alkyl moiety in which the methylene group is substituted with NR ^A , where R ^A is hydrogen, substituted or unsubstituted (C ₁ -C ₆ )alkyl, substituted or unsubstituted is selected from the group consisting of substituted (C ₃ -C ₈ )cycloalkyl, substituted or unsubstituted (C ₆ -C ₁₄ )aryl and substituted or unsubstituted (C ₃ -C ₁₄ )heteroaryl; In some embodiments, heteroalkyl is -CH ₂ SCH ₃ . In some embodiments, heteroalkyl is -CH ₂ OCH ₃ .

As used herein and throughout the description, the term “alkenyl” refers to an unsaturated straight or branched chain hydrocarbon monovalent radical having at least one carbon-carbon double bond. Generally, the maximum number of carbon-carbon double bonds in an alkenyl group is determined by dividing the number of carbon atoms in the alkenyl group by 2 (if the number of carbon atoms in the alkenyl group is odd, the result of the division is rounded down to an integer). For example, for an alkenyl group having nine carbon atoms, the maximum number of carbon-carbon double bonds is four. Preferably, alkenyl groups have 1 to 4, ie 1, 2, 3 or 4, carbon-carbon double bonds. Preferably, alkenyl groups have 2 to 10 carbon atoms, i.e. 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, more preferably 2 to 8 carbon atoms, For example, it contains 2 to 6 carbon atoms or 2 to 4 carbon atoms. Thus, in preferred embodiments, the alkenyl group contains 2 to 10 carbon atoms and 1, 2, 3, 4 or 5 carbon-carbon double bonds, more preferably 2 to 8 carbon atoms and 1, 2, 3 or 4 carbon-carbon double bonds, such as 2-6 carbon atoms and 1, 2 or 3 carbon-carbon double bonds, or 2-4 carbon atoms and 1 or It contains two carbon-carbon double bonds. In certain embodiments, the alkenyl group employed in the invention contains 2-20 carbon atoms (C _2-20 alkenyl). In some embodiments, the alkenyl group employed in the invention contains 2-15 carbon atoms (C _2-15 alkenyl). In another embodiment, the alkenyl group employed contains _2-10 carbon atoms (C _2-10 alkenyl). In still other embodiments, the alkenyl group contains _2-8 carbon atoms (C _2-8 alkenyl). In still other embodiments, the alkenyl group contains 2-6 carbons (C _2-6 alkenyl). In still other embodiments, the alkenyl group contains 2-5 carbons (C _2-5 alkenyl). In still other embodiments, the alkenyl group contains 2-4 carbons (C _2-4 alkenyl). In still other embodiments, the alkenyl group contains 2-3 carbons (C _2-3 alkenyl). In still other embodiments, the alkenyl group contains ₂ carbons (C2 alkenyl). Carbon-carbon double bonds may be in the cis (Z) or trans (E) configuration. Exemplary alkenyl groups are vinyl, 1-propenyl, 2-propenyl (ie, allyl), 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl, 1-octenyl, 2- octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6-octenyl, 7-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 4-nonenyl, 5-nonenyl, 6-nonenyl, 7-nonenyl, Including 8-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl, 4-decenyl, 5-decenyl, 6-decenyl, 7-decenyl, 8-decenyl, 9-decenyl and the like. If the alkenyl group is attached to a nitrogen atom, the double bond cannot be alpha to the nitrogen atom. Alkenyl group substituents include, but are not limited to, any of the substituents described herein that result in the formation of stable moieties. Substituted alkenyl groups can be substituted at any position so long as the resulting compound is sufficiently stable and suitable as a pharmaceutically active compound. The prerequisite that certain groups and compounds of any one of formulas I, II, III and IV are sufficiently stable and suitable as pharmaceutically active compounds is generally any of formulas I, II, III and IV Applies to all groups of a compound. In some embodiments, the alkenyl group is selected from halogen, hydroxyl, cyano, (C ₁ -C ₆ )alkyl-O- and (C ₁ -C ₆ )alkyl-S(O) _m -, 1 It may be substituted by one or more same or different substituents. In one embodiment of the invention, the numbers m are selected from 0 and 2 and all numbers m are independent of each other and may be the same or different. In some embodiments, the alkenyl chain is straight. In some embodiments the alkenyl chain is branched. In some embodiments the alkenyl chain is substituted. In some embodiments the alkenyl chain is unsubstituted. In some embodiments, alkenyl chains are linear and substituted or unsubstituted. In some embodiments, alkenyl chains are branched and substituted or unsubstituted. Alkenyl group substituents include, but are not limited to, any of the substituents described herein that result in the formation of stable moieties.

As used herein, throughout the description the terms "cycloalkyl" or "alicyclic" or "carbocyclic" or "carbocycle" preferably have 3 to 8 carbon atoms, i.e. 3 , 4, 5, 6, 7 or 8 carbon atoms, more preferably 3-7 carbon atoms, and cyclic non-aromatic versions of "alkyl" and "alkenyl". In another embodiment, the cycloalkyl group employed in the invention contains 3-7 carbon atoms (C _3-7 cycloalkyl). In another embodiment, the cycloalkyl group employed in the invention contains 3-6 carbon atoms (C _3-6 cycloalkyl). In another embodiment, the cycloalkyl group employed in the invention contains 3-5 carbon atoms (C _3-5 cycloalkyl). In another embodiment, the cycloalkyl group employed in the invention contains 3-4 carbon atoms (C _3-4 cycloalkyl). In another embodiment, the cycloalkyl group employed in the invention contains ₃ carbon atoms (C3cycloalkyl). Exemplary cycloalkyl groups include cyclopropyl, cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl and cyclooctenyl. The double bond in the cycloalkenyl group is to the carbon atom in the 1-position (the nitrogen atom through which the group is attached to the rest of the molecule, i.e., for example, the nitrogen atom in compounds having a structure according to Formula I). It can be in any position, thus cycloalkenyl is for example cyclopent-1-enyl, cyclopent-2-enyl, cyclopent-3-enyl, cyclohex-1-enyl, cyclohex-2-enyl, cyclohex-3-enyl , cyclohept-1-enyl, cyclohept-2-enyl, cyclohept-3-enyl, cyclohept-4-enyl. In a preferred embodiment of the invention, a cycloalkyl group such as (C ₃ -C ₇ )-cycloalkyl in the definition of any group refers to any two or more subgroups of said specified cycloalkyl groups, for example , from cyclopropyl and cyclobutyl, or from cyclopropyl, cyclobutyl and cyclopentyl, or from cyclopropyl, cyclopentyl and cyclohexyl, or from cyclopentyl and cyclohexyl, from cyclopentyl, cyclohexyl and cycloheptyl. Similarly, in preferred embodiments, the cycloalkenyl group may be from two or more subgroups of any of the above specified cycloalkenyl groups, such as from cyclopentenyl and cyclohexenyl, or from cyclohexenyl and cycloheptenyl, or from cyclopent-1- from enyl, cyclopent-2-enyl, cyclohex-1-enyl, cyclohex-2-enyl, cyclohept-1-enyl and cyclohept-2-enyl, or cyclopent-2-enyl, cyclopent-3-enyl, cyclohex-2-enyl from enyl, cyclohex-3-enyl, cyclohept-2-enyl, cyclohept-3-enyl and cyclohept-4-enyl, or from cyclopent-2-enyl and cyclohex-2-enyl, or from cyclopent-2-enyl, cyclohexa- is selected from 2-enyl and cyclohept-2-enyl; Cycloalkyl and cycloalkenyl groups are generally optionally substituted with one or more (C ₁ -C ₄ )-alkyl substituents. That is, they are unsubstituted, i.e. have no alkyl substituents, or may be present in any position, for example 1, 2, 3 or 4 identical or different (C ₁ -C ₄ )-substituted by alkyl substituents such as methyl groups and/or ethyl groups and/or isopropyl groups and/or tert-butyl groups, especially methyl groups. Examples of alkyl-substituted cycloalkyl groups are 1-methyl-cyclopropyl, 2,2-dimethyl-cyclopropyl, 1-methyl-cyclopentyl, 2,3-dimethyl-cyclopentyl, 1-methyl-cyclohexyl, 4-methyl-cyclohexyl. , 4-isopropyl-cyclohexyl, 4-tert-butyl-cyclohexyl and 3,3,5,5-tetramethyl-cyclohexyl. Examples of alkyl-substituted cycloalkenyl groups are 1-methyl-cyclopent-2-enyl, 2-methyl-cyclopent-2-enyl, 3-methyl-cyclopent-2-enyl, 3,4-dimethyl-cyclopent-3-enyl. , 1-methyl-cyclohex-2-enyl, 2-methyl-cyclohex-2-enyl, 3-methyl-cyclohex-2-enyl, 4-methyl-cyclohex-2-enyl, 2-methyl-cyclohex-3-enyl , 3-methyl-cyclohex-3-enyl, 4-methyl-cyclohex-3-enyl, 2,3-dimethyl-cyclohex-2-enyl, 4,4-dimethyl-cyclohex-2-enyl, 3,4-dimethyl -cyclohex-3-enyl. Cycloalkyl and cycloalkenyl groups are also typically optionally substituted with one or more fluorine atoms. ie they are unsubstituted, i.e. they have no fluorine atoms, or they have, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 fluorine atoms, preferably substituted by 1, 2, 3, 4, 5 or 6 fluorine atoms. Cycloalkyl and cycloalkenyl groups can also be substituted simultaneously with fluorine and alkyl. Fluorine atoms can be present in any position and can also be present in the alkyl substituents. Examples of fluoro-substituted cycloalkyl groups are 1-fluoro-cyclopropyl, 2,2-difluoro-cyclopropyl, 3,3-difluoro-cyclobutyl, 1-fluoro-cyclohexyl, 4,4-difluoro-cyclohexyl and 3,3 , 4,4,5,5-hexafluoro-cyclohexyl. Examples of fluoro-substituted cycloalkenyl groups are 1-fluoro-cyclopent-2-enyl, 1-fluoro-cyclohex-2-enyl, 4-fluoro-cyclohex-2-enyl, 4,4-difluoro-cyclohex-2-enyl. is. In one embodiment of the invention, the cycloalkyl group is optionally unsubstituted by substituents selected from fluorine and (C ₁ -C ₄ )-alkyl. When a cycloalkyl or cycloalkenyl group may be substituted by further substituents such as hydroxy, it may only be substituted by one or more further substituents such as hydroxy, and fluorine and (C ₁ -C ₄ )-alkyl, or by one or more further substituents, etc. and at the same time one or more substituents selected from fluorine and (C ₁ -C ₄ )-alkyl It can be substituted by substituents. The number of such further substituents such as hydroxy which may be present on a cycloalkyl or cycloalkenyl group is preferably 1, 2 or 3, more preferably 1 or 2, eg 1. The total number of all substituents in a cycloalkyl or cycloalkenyl group is preferably 1, 2, 3, 4, 5, 6, 7 or 8, more preferably 1, 2, 3, 4 or 5, such as 1 , 2 or 3. Such additional substituents, such as hydroxy, can be present at any position as long as the resulting compound is sufficiently stable and suitable as a subgroup in pharmaceutically active compounds. Preferably, no hydroxy substituent is present at the 1-position of a cycloalkenyl or cycloalkyl group, and in cycloalkenyl groups the hydroxy substituent is not present on a carbon atom that is part of a double bond. Examples of hydroxy-substituted cycloalkyl groups are 3-hydroxy-cyclobutyl, 2-hydroxy-cyclopentyl, 3-hydroxy-cyclopentyl, 3,4-dihydroxy-cyclopentyl, 2-hydroxy-cyclohexyl, 3-hydroxy-cyclohexyl, 4-hydroxy -cyclohexyl, 2,3-dihydroxy-cyclohexyl, 2,4-dihydroxy-cyclohexyl, 3,4-dihydroxy-cyclohexyl, 3,5-dihydroxy-cyclohexyl, 3,4,5-trihydroxy-cyclohexyl, 2-hydroxy- Cycloheptyl, 3-hydroxy-cycloheptyl, 4-hydroxy-cycloheptyl. Examples of hydroxy-substituted cycloalkenyl groups are 5-hydroxy-cyclopent-2-enyl, 4-hydroxy-cyclohex-2-enyl, 5-hydroxy-cyclohex-2-enyl, 6-hydroxy-cyclohex-2-enyl, 6 -hydroxy-cyclohex-3-enyl. The term "cycloalkyl" is also meant to include bicyclic and tricyclic versions thereof. When a bicyclic ring is formed, each ring is preferably linked together at two adjacent carbon atoms, but alternatively the two rings are linked via the same carbon atom, i.e. they forms a spiro ring system or forms a "bridged" ring system. Preferred examples of cycloalkyl are C ₃ -C ₈ -cycloalkyl, especially cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, spiro[3,3]heptyl, spiro[3,4]octyl, spiro[ 4,3]octyl, bicyclo[4.1.0]heptyl, bicyclo[3.2.0]heptyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[5. 1.0]octyl and bicyclo[4.2.0]octyl. Cycloalkyl group substituents include, but are not limited to, any of the substituents described herein that result in the formation of stable moieties.

As used herein, throughout the description, the term "cyclopropylene" means a cyclopropyl group, as defined above, with one hydrogen atom removed to yield a divalent group. Cyclopropylene links two atoms or moieties via the same carbon atom (1,1-cyclopropylene, ie a geminal divalent group) or via two carbon atoms (1,2-cyclopropylene) can do.

As used herein, the term “aryl” or “aromatic ring” throughout the description refers to an aromatic monocyclic or polycyclic ring system having from 6 to 10 ring atoms, the ring atoms of which All are carbon and may be substituted or unsubstituted. _In certain embodiments of the invention, "aryl" refers to a monocyclic or bicyclic C6 _- C10 aromatic ring system having one or two aromatic rings, including phenyl, biphenyl, naphthyl, and the like. including but not limited to, may have one or more substituents. Preferably, aryl groups contain 6-10 carbon atoms, which can be arranged in a single ring (eg, phenyl) or two or more condensed rings (eg, naphthyl). Exemplary aryl groups include cyclopropenylium, cyclopentadienyl, phenyl, indenyl, naphthyl and azulenyl. Preferably, "aryl" refers to a monocyclic ring containing 6 carbon atoms or an aromatic bicyclic ring system containing 10 carbon atoms. Preferred examples are substituted or unsubstituted phenyl and naphthyl. Even more preferred is substituted or unsubstituted phenyl. In certain embodiments, the aryl group employed in the invention contains 7-10 carbon atoms (C _7-10 aryl). In another embodiment, the aryl group employed in the invention contains 6-8 carbon atoms (C _6-8 aryl). In another embodiment, the aryl group employed in the invention contains ₆ carbon atoms (C6 aryl). In another embodiment, the aryl group employed in the invention contains ₁₀ carbon atoms (C10 aryl).

In substituted aryl groups, substituents can be present at any position. In monosubstituted phenyl groups the substituent can be in the 2-, 3- or 4-position. In disubstituted phenyl groups the substituents can be in the 2,3-position, 2,4-position, 2,5-position, 2,6-position, 3,4-position or 3,5-position. . In trisubstituted phenyl groups, the substituents may be in 2,3,4-positions, 2,3,5-positions, 2,3,6-positions, 2,4,5-positions, 2,4,6-positions or 3-positions. , 4,5-position. When a phenyl group has 4 substituents, 1, 2, 3 or 4 of which can be fluorine atoms, for example, the unsubstituted ring carbon atoms are in the 2-position, the 3-position or may be in the 4-position. When a polysubstituted phenyl group or heteroaryl group has different substituents, each substituent can be present in any suitable position and the present invention includes all positional isomers. The number of substituents in a substituted phenyl group can be 1, 2, 3, 4 or 5. Preferably, substituted phenyl groups, and likewise substituted heteroaryl groups, have 1, 2 or 3, especially 1 or 2, identical or different substituents. In preferred embodiments, substituted phenyl groups have one substituent at the 2, 3 or 4 positions. In preferred embodiments of the invention, substituents in substituted phenyl groups and substituted heteroaryl groups are selected from any subgroup of substituents recited in each definition, such as halogen, substituted or unsubstituted ( C ₁ -C ₆ )-alkyl, (C ₁ -C ₄ )-haloalkyl, —NO ₂ , cyano, halogen, substituted or unsubstituted (C ₁ -C ₆ )alkyl-O—, substituted or unsubstituted (C ₆ —C ₁₄ )aryl(C ₁ -C ₆ )alkyl-O—, —C(O)NH ₂ , —C(O)NH((C ₁ -C ₆ )-alkyl), —C(O)N( (C ₁ -C ₆ )-alkyl) ₂ , —NH((C ₁ -C ₆ )alkyl-O(C ₁ -C ₆ )alkyl)), —NH((C ₁ -C ₆ )alkyl-OH) , azide, —NH(C(=O)(C ₁ -C ₆ )alkyl) and —N((C ₁ -C ₆ )alkyl)(C(=O)(C ₁ -C ₆ )alkyl) depending on the substituents used.

As used herein and throughout the description, the term "arylene" refers to an aryl divalent group derived by the removal of two hydrogen atoms from an aryl group as defined herein. Arylene groups can be substituted or unsubstituted. Arylene group substituents include, but are not limited to, any of the substituents described herein that result in the formation of stable moieties.

As used herein, throughout the description, the term "heteroaryl" or "heteroaromatic ring" means that one or more carbon atoms in the aryl group are replaced with O, S or N heteroatoms. aryl group as defined above. Preferably, the heteroaryl group contains 3-10 carbon atoms. Preferably, heteroaryl refers to a 5- or 6-membered aromatic monocyclic ring in which 1, 2 or 3 carbon atoms are replaced with the same or different heteroatoms of O, N or S. Alternatively, it means an aromatic bicyclic or tricyclic ring system in which 1, 2, 3, 4 or 5 carbon atoms are replaced with the same or different heteroatoms of O, N or S. Preferably, the maximum number of O atoms is 1, the maximum number of S atoms is 1 and the maximum total number of O and S atoms is 2 in each ring of the heteroaryl group. In certain embodiments, the heteroaryl groups employed in the invention are 5-membered aromatic monocyclic rings in which 1, 2 or 3 carbon atoms are replaced by the same or different heteroatoms of O, N, or S. It is a ring. In certain embodiments, the heteroaryl groups employed in the invention are 5-membered aromatic monocyclic rings in which 1, 2 or 3 carbon atoms have been replaced by the same or different O heteroatoms. In certain embodiments, the heteroaryl group employed in the invention is a 5-membered aromatic monocyclic ring having 1, 2 or 3 carbon atoms replaced by the same or different heteroatoms of O and N. is. In certain embodiments, the heteroaryl group employed in the invention is a 5-membered aromatic monocyclic ring in which 1, 2 or 3 carbon atoms are replaced by the same or different heteroatoms of O and S. be. In certain embodiments, the heteroaryl group employed in the invention is a 5-membered aromatic monocyclic ring in which 1, 2 or 3 carbon atoms are replaced by the same or different heteroatoms of N and S. be. In certain embodiments, the heteroaryl group employed in the invention is a 6-membered aromatic group in which 1, 2 or 3 carbon atoms are replaced by the same or different heteroatoms of O, S or N. It is a monocyclic ring. In certain embodiments, the heteroaryl group employed in the invention is a 6-membered aromatic monocyclic ring in which 1, 2 or 3 carbon atoms are replaced by N. In certain embodiments, the heteroaryl groups employed in the invention are aromatic divalent groups in which 1, 2, 3, 4 or 5 carbon atoms have been replaced by the same or different heteroatoms of O, N or S. It is a cyclic system. Exemplary heteroaryl groups include furanyl, thienyl, oxazolyl, isoxazolyl, oxadiazolyl (1,2,5- and 1,2,3-), pyrrolyl, imidazolyl, pyrazolyl, triazolyl (1,2,3- and 1, 2,4-), tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl (1,2,3- and 1,2,5-), pyridyl, pyrimidinyl, pyrazinyl, triazinyl (1,2,3-, 1,2,4- and 1,3,5-), benzofuranyl (1- and 2-), indolyl, azaindolyl (4-, 5-, 6- and 7-), diazaindolyl, isoindolyl, benzothienyl (1- and 2- -), 1H-indazolyl, benzimidazolyl, benzoxazolyl, indoxazinyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzotriazolyl, quinolinyl, isoquinolinyl, benzodiazinyl, quinoxalinyl, quinazolinyl, benzotriazinyl (1,2) ,3- and 1,2,4-benzotriazinyl), pyridazinyl, phenoxazinyl, thiazolopyridinyl, pyrrolotiazolyl, phenothiazinyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathiinyl, pyrrolidinyl, indolizinyl, indazolyl , purinyl, quinolidinyl, phthalazinyl, naphthyridinyl (1,5-, 1,6-, 1,7-, 1,8- and 2,6-), cinolinyl, pteridinyl, carbazolyl, phenanthridinyl, acridinyl, perimidinyl, Phenanthrolinyl (1,7-, 1,8-, 1,10-, 3,8- and 4,7-), phenazinyl, oxazolopyridinyl, isoxazolopyrodinyl, pyrrolooxazolyl , pyrrolopyrrolyl and the like, which may have one or more substituents. Exemplary 5- or 6-membered heteroaryl groups include furanyl, thienyl, oxazolyl, isoxazolyl, oxadiazolyl (1,2,5- and 1,2,3-), pyrrolyl, imidazolyl, pyrazolyl, triazolyl (1,2, 3- and 1,2,4-), thiazolyl, isothiazolyl, thiadiazolyl (1,2,3- and 1,2,5-), pyridyl, pyrimidinyl, pyrazinyl, thiazinyl (1,2,3-, 1,2 ,4- and 1,3,5-) and pyridazinyl. Exemplary bicyclic heteroaryl groups include 7-azaindolyl, 6-azaindolyl, 5-azaindolyl, 4-azaindolyl, benzofuranyl and indolyl. Heteroaryl substituents include, but are not limited to, any of the substituents described herein that result in the formation of stable moieties.

In substituted heteroaryl groups, the substituents can be at any position, for example, thiophen-3- at the 2- and/or 4- and/or 5-position of the pyridin-2-yl group, at the 3- and/or 4- and/or 5-position of the pyridin-2-yl group and /or at the 6-position, at the 2- and/or 4- and/or 5- and/or 6-position of the pyridin-3-yl group, at the 2-position and/or of the pyridin-4-yl group 1H-pyrazol-1-yl at the 3-position and/or the 5-position and/or the 6-position, at the 3-position and/or the 5-position and/or the 1-position of the 1H-pyrazol-4-yl group in the 3- and/or 5- and/or 4-position of the group, in the 2- and/or 3- and/or 4- and/or 5-position of the 1H-indol-1-yl group and /or at the 6- and/or 7-position, at the 1- and/or 2- and/or 4- and/or 5- and/or 6- and/or 7-positions of the 1H-indol-3-yl group can exist. A substituted heteroaryl group may be monosubstituted or polysubstituted, ie, having more than one substituent. In preferred embodiments, the 1H-indol-1-yl group is substituted in the 2- and 3-positions and/or in the 6- and 7-positions. In other preferred embodiments, the 1H-indol-3-yl group is substituted in the 1 and 7 positions and/or in the 6 and 7 positions. Preferably, a substituted heteroaryl group is substituted by 1, 2 or 3, especially 1 or 2, for example 1, identical or different substituents. When a hydrogen atom or a ring nitrogen atom capable of bearing a substituent is present, the substituent on this nitrogen atom can be, for example, a methyl, ethyl, propyl or t-butyl group, these groups also It can be monosubstituted or polysubstituted by fluorine. In general, a suitable ring nitrogen atom of the aromatic ring of a heteroaryl group, such as the nitrogen atom of a pyridinyl group, may also carry an oxide substituent ^—O— , thereby providing compounds of Formula I, Formula II, Formula III and Formula IV. exists in the form of the N-oxide.

As used herein and throughout the description, the term "heteroarylene" refers to a divalent radical derived from a heteroaryl group as defined herein by the removal of two hydrogen atoms. A heteroarylene group can be substituted or unsubstituted.

As used herein, throughout the description the terms "arylalkyl" and "heteroarylalkyl" refer to groups in which the aryl and heteroaryl groups, respectively, are attached to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, etc.) (including alkyl groups in which a carbon atom (e.g., methylene group) is substituted, for example, by an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, etc. )). Preferably, arylalkyl is substituted or unsubstituted (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl. Preferably, heteroarylalkyl is substituted or unsubstituted (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl. In some embodiments, the alkyl chain is straight. In some embodiments, the alkyl chain is branched. In some embodiments, the alkyl chain is substituted. In some embodiments, alkyl chains are unsubstituted. In some embodiments, alkyl chains are linear and substituted or unsubstituted. In some embodiments, alkyl chains are branched and substituted or unsubstituted.

As used herein, throughout the description, the terms "arylheteroalkyl" and "heteroarylheteroalkyl" are meant to include groups in which aryl and heteroaryl groups, respectively, are attached to a heteroalkyl group. do. In some embodiments, the heteroalkyl chain is straight. In some embodiments, heteroalkyl chains are branched. In some embodiments, the heteroalkyl chain is substituted. In some embodiments, the heteroalkyl chain is unsubstituted. In some embodiments, the heteroalkyl chain is linear and substituted or unsubstituted. In some embodiments, heteroalkyl chains are branched, substituted or unsubstituted.

As used herein and throughout the description, the term "halogen" or "halo" means fluoro, chloro, bromo or iodo.

As used herein, the term "cyano" means -CN throughout the description.

As used herein _, the term "azido" means N3 throughout the description.

As used herein, the term “optionally substituted” or “substituted” throughout the description refers to one or more (eg, from 1 to the maximum number of hydrogen atoms attached to a group, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or up to 10, for example between 1 to 5, 1 to 4 or 1 to 3, or 1 or 2) hydrogen atoms are hydrogen different groups ((C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )heteroalkyl, (C ₁ -C ₆ )haloalkyl, (C ₂ -C ₆ )alkenyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ ) alkyl, halogen, —CN, —NO ₂ , —OR ¹⁶¹ , —N(R ¹⁶² )(R ¹⁶³ ), —N(R ¹⁶¹ )(OR ¹⁶¹ ), —S(O) _0-2 R ¹⁶¹ , —S(O) _1-2 OR ¹⁶¹ , —OS(O) _1-2 R ¹⁶¹ , —OS(O) _1-2 OR ¹⁶¹ , —S(O) _1-2 N(R ¹⁶² )(R ¹⁶³ ) , —OS(O) _1-2 N(R ¹⁶² )(R ¹⁶³ ), —N(R ¹⁶¹ )S(O) _1-2 R ¹⁶¹ , —NR ¹⁶¹ S(O) _1-2 OR ¹⁶¹ , —NR ¹⁶¹ S(O) _1-2 N(R ¹⁶² )(R ¹⁶³ ), -C(=W)R ¹⁶¹ , -C(=W)WR ¹⁶¹ , -WC(=W)R ¹⁶¹ and -WC(=W ) WR ¹⁶¹ , etc. (R ¹⁶¹ , R ¹⁶² and R ¹⁶³ are independently selected from the group consisting of —H, alkyl, alkenyl, alkynyl, cycloalkyl, aryl and heteroaryl, preferably R ¹⁶¹ , R ¹⁶² and R ¹⁶³ is —H, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, 3-7 membered cycloalkyl, 5-6 membered aryl, 5-6 membered heteroaryl and 3-7 membered heterocyclyl R ¹⁶⁴ is independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and —OR ¹⁶¹ ; and W is O, S and —N(independently selected from R ¹⁶⁴ ))).

As used herein, throughout the description, the term "optionally" or "optionally" as used herein means whether or not the subsequently described event, circumstance or condition occurs. Often, description is meant to include instances in which the event, situation or condition occurs and instances in which it does not occur.

As used herein, throughout the description, the term "solvate" as used herein refers to solvents such as organic solvents such as aliphatic alcohols such as methanol, ethanol, n-propanol. , isopropanol, etc.), acetone, acetonitrile, ether, etc.), water, or a mixture of two or more of these liquids), where the adduct is a crystal or mixed crystal Exists in form. The amount of solvent included in the adduct complex may be stoichiometric or non-stoichiometric. A "hydrate" is a solvate in which the solvent is water.

As used herein and throughout the description, the term "pharmaceutically acceptable salt" refers to salts that retain the desired biological activity of the parent compound and do not impart any undesired toxicological effects. (see, eg, Berge, SM, et al. (1977) J. Pharm. Sci. 66: 1-19). Physiologically acceptable salts of the compounds of the present invention are especially salts with non-toxic salt components, preferably pharmaceutically acceptable salts. They can contain inorganic or organic salt components. Such salts are, for example, from compounds of the invention containing an acidic group, such as a carboxylic acid group (HO--CO--) or a sulfonic acid group (HO--S(O) ₂ --) and a non-toxic inorganic or organic base. can be formed. Suitable bases are, for example, alkali metal or alkaline earth metal compounds, such as sodium hydroxide, potassium hydroxide, sodium carbonate or sodium hydrogen carbonate, or ammonia, organic amino compounds and quaternary ammonium hydroxides. The reaction of a compound of the invention and a base for salt preparation is generally carried out in a solvent or diluent according to conventional methods. For physiological and chemical stability, advantageous salts of acidic groups are often sodium, potassium, magnesium or calcium salts or ammonium salts (which also carry one or more organic groups on the nitrogen atom). can be done). Compounds of the invention containing a basic, i.e. protonatable group, such as an amino group or another basic heterocyclic ring, can be added in the form of their acid addition salts with physiologically acceptable acids, for example hydrogen chloride , hydrogen bromide, phosphoric acid, sulfuric acid, acetic acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic acid, which are generally dissolved with acids in solvents or diluents according to conventional methods. can be prepared from the compounds of the present invention by reaction. As usual, particularly in the case of acid addition salts of compounds containing two or more basic groups, in the resulting salt the ratio of the salt components can deviate upwards or downwards from the stoichiometric ratio. For example, in the case of acid addition salts with monovalent acids of compounds of the invention containing 1 or 2 basic groups, a molar ratio of 1:1 or 1:2, which applies It can vary depending on the conditions.
The present invention also includes salts containing components in non-stoichiometric ratios, for example, that an acid addition salt of a compound of the invention is shown to contain equimolar amounts of an acid, so that in the resulting salt Lesser or greater amounts of acid are also contemplated, such as about 0.8 or about 1.1 moles of acid per mole of compounds of the invention. In addition to the salt forms mentioned, the invention also includes internal salts (betaines, zwitterions) when the compounds according to the invention contain simultaneously acidic and basic groups in the molecule. The present invention is also useful for chemical reactions or for the preparation of physiologically acceptable salts (e.g. anion exchange or including all salts of the compounds of the invention which are suitable as intermediates (by cation exchange). A subject of the invention is also the solvates of the compounds according to the invention and their salts, for example hydrates and adducts with alcohols such as (C ₁ -C ₄ )-alkanols, in particular physiologically acceptable are solvates as well as active metabolites of the compounds of the present invention.

As used herein, throughout the description, the term "pharmaceutically acceptable" is specifically for use in animals, more particularly in humans, approved by a regulatory agency or other generally recognized pharmacopoeia. It can mean that

The present invention includes all stereoisomers, including all possible enantiomers and diastereomers, including cis/trans isomers, of the compounds of Formula I. Likewise, the invention includes mixtures of two or more stereoisomers, eg mixtures of enantiomers and/or diastereomers, including cis/trans isomers, in all ratios.

All asymmetric centers contained in compounds of Formula I (for example, in unsubstituted or substituted alkyl groups or stereogenic carbons CH-B (when p and q are unequal) shown in Formula I) are It can independently have the S configuration or the R configuration. The present invention provides two compounds in both levorotatory and dextrorotatory enantiomers, in enantiomerically pure and substantially enantiomerically pure forms, and in racemic form, and in all ratios. It relates to enantiomers in the form of mixtures of enantiomers. The invention likewise relates to diastereomers in the form of pure and substantially pure diastereomers, as well as mixtures of two or more diastereomers in all ratios. The present invention also provides all cis/trans isomers of the compounds of formula I, for example in pure and substantially pure form, as well as mixtures of cis and trans isomers in all ratios. Contain in the form. Cis/trans isomerism can occur, for example, in substituted cycloalkane rings. The preparation of individual stereoisomers, if desired, can be carried out by conventional methods, such as resolution of mixtures by chromatography or crystallization, or by the use of stereochemically uniform starting compounds in the synthesis, or by stereoselective reactions. can be done by Optionally, derivatization can be carried out prior to separation of stereoisomers. Separation of mixtures of stereoisomers can be carried out at the stage of compounds of formula I or at intermediate stages in the course of the synthesis. The invention also includes all tautomeric forms of the compounds of the invention.

（式中、
Ｘは、Ｎ及びＣＨからなる群より選択され；
Ｒ^１は、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル、（Ｃ_３－Ｃ_８）シクロアルキル、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール及び（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
Ｒ^２及びＲ^３は、独立して、水素、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）ハロアルキル及び（Ｃ_３－Ｃ_８）シクロアルキルからなる群より選択され；
ｎは、０と３の間の整数であり；
Ｚは、Ｃ＝Ｏ、Ｃ＝Ｓ及びＣＨ_２からなる群より選択され；
Ａは、－Ｎ（Ｒ^４）（Ｒ^５）及び下記式：

の基からなる群より選択され；
Ｒ^４は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^５は、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）ヘテロアルキル及び（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）ヘテロアルキルからなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^６は、（Ｃ_１－Ｃ_４）アルキル又は（Ｃ_１－Ｃ_４）ハロアルキルであり；
Ｙは、Ｎ－Ｂ、ＣＨ－Ｂ及びＯからなる群より選択され；
Ｂは、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）アルキル及びＮ（Ｒ^７）（Ｒ^８）からなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^７は、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^８は、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
ｐは、１と２の間の整数であり、
ｑは、１と３の間の整数である）
による構造を有する化合物であって、その立体異性型のいずれか、又は任意の比率の立体異性型の混合物であるもの、ならびに、その生理学的に許容される塩、及びそれらのいずれかの生理学的に許容される溶媒和物又は水和物を提供する。 In one aspect, the present invention provides a compound of formula I:

(In the formula,
X is selected from the group consisting of N and CH;
R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) selected from the group consisting of aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl;
R ² and R ³ are independently selected from the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl;
n is an integer between 0 and 3;
Z is selected from the group consisting of C=O, C=S and _CH2 ;
A is —N(R ⁴ )(R ⁵ ) and the following formula:

selected from the group consisting of
R ⁴ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ ;
R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl;
Y is selected from the group consisting of NB, CH-B and O;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl and N(R ⁷ )(R ⁸ ) which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ ;
R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl ( selected from the group consisting of C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl;
p is an integer between 1 and 2;
q is an integer between 1 and 3)
in any of its stereoisomeric forms or mixtures of stereoisomeric forms in any ratio, and physiologically acceptable salts thereof, and physiologically acceptable salts of any of them. to provide an acceptable solvate or hydrate for

In some embodiments, X is N. In some embodiments, X is _CH2 .

In some embodiments, R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, ( selected from the group consisting of C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl; In other embodiments, hydrogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl, (C ₃ -C ₆ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) aryl(C ₁ -C ₄ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₄ )alkyl, in other embodiments hydrogen, ( C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ ) alkyl, in other embodiments hydrogen, (C ₁ -C ₆ )alkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, in other embodiments hydrogen, (C ₁ -C ₄ )alkyl, (C ₃ -C ₆ )cycloalkyl, (C ₆ -C ₁₀ )aryl and (C ₆ -C ₁₀ ) aryl(C ₁ -C ₄ )alkyl, in other embodiments hydrogen, (C ₁ -C ₄ )alkyl, (C ₃ -C ₆ )cycloalkyl, (C ₆ )aryl and (C ₆ )aryl from (C ₁ -C ₄ )alkyl, in other embodiments hydrogen, from (C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )haloalkyl, in other embodiments hydrogen, (C ₁ -C ₄ ) from alkyl and (C ₁ -C ₄ )haloalkyl, in other embodiments hydrogen, from (C ₂ -C ₆ )alkyl and (C ₂ -C ₆ )haloalkyl, in other embodiments hydrogen, ( from C ₃ -C ₆ )alkyl and (C ₃ -C ₆ )haloalkyl, in other embodiments hydrogen and (C1-C6) selected from alkyl, in other embodiments from hydrogen and (C ₁ -C ₄ )alkyl, in other embodiments from the group consisting of hydrogen and (C ₃ -C ₄ )alkyl; R ¹ is (C ₁ -C ₆ )alkyl, preferably (C ₁ -C ₄ )alkyl, more preferably (C ₁ -C ₂ )alkyl, even more preferably methyl. In some embodiments, R ¹ is (C ₆ -C ₁₀ )aryl, preferably wherein aryl is selected from the group of substituted or unsubstituted naphthyl and substituted or unsubstituted phenyl, and more Preferably, aryl here is substituted or unsubstituted phenyl. In some embodiments, R ¹ is (C ₁ -C ₆ )haloalkyl, preferably —CF ₃ , and in other embodiments (C ₃ -C ₈ )cycloalkyl, preferably substituted or unsubstituted is cyclopropyl, and in another embodiment (C ₆ -C ₁₀ )aryl(C ₁ -C 6 )alkyl, preferably (C ₆ )aryl(C ₁ -C ₆ )alkyl, more preferably substituted or unsubstituted is the benzyl of Alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl are substituted or unsubstituted (C ₁ -C ₆ )alkyl, substituted or unsubstituted (C ₁ -C ₆ )haloalkyl, —OR ²⁶ , halogen and cyano, preferably substituted with one or more identical or different substituents selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ²⁶ also, wherein R ²⁶ is hydrogen, substituted or unsubstituted (C ₁ -C ₆ )alkyl and substituted or unsubstituted (C ₂ -C ₆ )alkenyl, preferably hydrogen and substituted or unsubstituted (C ₁ -C ₆ ) selected from the group consisting of alkyl; Preferably the number of substituents in a substituted R ¹ group is 1, 2, 3 or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents in substituted R ¹ groups are present on any position of the carbon atom as generally described above for substituted alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl groups. be able to.

In some embodiments, R ² and R ³ are selected from the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl; In other embodiments hydrogen, (C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )haloalkyl, in other embodiments (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl, in other embodiments independently selected from the group consisting of (C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )haloalkyl, in other embodiments , R ² and R ³ are hydrogen, and in other embodiments, R ² and R ³ are (C ₁ -C ₆ )alkyl, preferably (C ₁ -C ₄ )alkyl, more preferably (C ₁ —C ₂ )alkyl, even more preferably methyl; in other embodiments, R ² and R ³ are (C ₁ -C ₆ )haloalkyl, preferably (C ₁ -C ₄ )haloalkyl, more preferably (C ₁ -C ₂ )haloalkyl, even more preferably —CF ₃ , in other embodiments one of R ² and R ³ is hydrogen and the other is (C ₁ -C ₆ )haloalkyl, preferably methyl, in other embodiments one of R ² and R ³ is hydrogen and the other is (C ₁ -C ₆ )haloalkyl, preferably —CF ₃ , in other embodiments R ² and One of R ³ is hydrogen and the other is (C ₃ -C ₈ )cycloalkyl, preferably cyclopropyl, in other embodiments one of R ² and R ³ is (C ₁ -C ₆ )alkyl, preferably (C ₁ -C ₄ )alkyl, more preferably (C ₁ -C ₂ )alkyl, even more preferably methyl, the other is (C ₁ -C ₆ )haloalkyl, preferably (C ₁ —C ₄ )haloalkyl, more preferably (C ₁ -C ₂ )haloalkyl, even more preferably —CF ₃ , in other embodiments one of R ² and R ³ is (C ₁ -C ₆ )alkyl; preferably (C ₁ -C ₄ )alkyl, more preferably (C ₁ -C ₂ )alkyl, even more preferably methyl, the other being (C ₃ -C ₈ )alkyl cycloalkyl, preferably (C ₃ -C ₆ )cycloalkyl, more preferably cycloalkyl; in another embodiment one of R ² and R ³ is methyl and the other is isopropyl or ethyl; In embodiments, one of R ² and R ³ is selected from the group consisting of methyl, ethyl, propyl and isopropyl and the other is (C ₁ -C ₆ )haloalkyl, preferably —CF ₃ , —CH ₂ CF ₃ , is selected from -CF ₂ CF ₃ , more preferably -CF ₃ ; Alkyl and cycloalkyl are substituted or unsubstituted (C ₁ -C ₆ )alkyl, substituted or unsubstituted (C ₁ -C ₆ )haloalkyl, —OR ²⁶ , halogen and cyano, preferably halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ²⁶ , wherein R ²⁶ is hydrogen, substituted or non- substituted (C ₁ -C ₆ )alkyl and substituted or unsubstituted (C ₂ -C ₆ )alkenyl, preferably selected from hydrogen and substituted or unsubstituted (C ₁ -C ₆ )alkyl. Preferably the number of substituents in the substituted ^R2 and/or R3 groups is 1, 2, ³ or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents in substituted ^R2 and/or R3 groups can be present ^on any position of the carbon atom, as generally described above for substituted alkyl and cycloalkyl groups.

In some embodiments n is an integer between 0 and 3, in other embodiments n is 0, 1, 2 or 3, in other embodiments n is between 1 and 3 and in other embodiments n is an integer between 1 and 2, in other embodiments n is 1, in other embodiments n is 2, in other embodiments In embodiments, n is 3, and in other embodiments n is 0.

In some embodiments Z is selected from the group consisting of C=O, C=S and _CH2 , in other embodiments C=O and C=S, in other embodiments C =O and _CH2 , in other embodiments C=S and _CH2 , in other embodiments Z is C=O, in other embodiments Z is C=S and in other embodiments, Z is _CH2 .

の基からなる群より選択され、他の実施形態において、－Ｎ（Ｒ^４）（Ｒ^５）及び下記式：

の基から選択され、他の実施形態において、下記式：

の基から選択され、他の実施形態において、－Ｎ（Ｒ^４）（Ｒ^５）であり、他の実施形態において、下記式：

の基であり、他の実施形態において、下記式：

の基であり、他の実施形態において、下記式：

の基であり、他の実施形態において、下記式：

の基である。 In some embodiments, A is —N(R ⁴ )(R ⁵ ) as well as the formula:

and in other embodiments -N(R ⁴ )(R ⁵ ) and the formula:

and in other embodiments, the formula:

in another embodiment -N(R ⁴ )(R ⁵ ), in another embodiment:

and in another embodiment, the formula:

and in another embodiment, the formula:

and in another embodiment, the formula:

is the basis of

In some embodiments, R ⁴ is hydrogen or (C ₁ -C ₆ )alkyl, in other embodiments hydrogen or (C ₁ -C ₄ )alkyl, in other embodiments hydrogen or (C ₃ -C ₄ )alkyl, in other embodiments hydrogen or (C ₁ -C ₂ )alkyl, in other embodiments R ⁴ is hydrogen, in other embodiments R ⁴ is (C ₁ -C ₆ )alkyl, preferably (C ₁ -C ₄ )alkyl, more preferably (C ₁ -C ₂ )alkyl, even more preferably methyl. Alkyl may be optionally substituted by one or more same or different substituents selected from the group consisting of halogen and cyano, preferably halogen. Preferably the number of substituents in a substituted ^R4 group is 1, 2, 3 or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents in substituted ^R4 groups can be present on any position of the carbon atom, as generally described above for substituted alkyl groups.

In some embodiments, R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ - (C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl; in another embodiment, (C ₆ )aryl (C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ ) From aryl(C ₁ -C ₆ )heteroalkyl, in other embodiments, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl, in other embodiments from (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl and (C ₃ - selected from C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably from (C ₆ )aryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl be done. In some embodiments, R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, preferably wherein aryl is substituted or unsubstituted naphthyl and substituted or unsubstituted phenyl more preferably wherein aryl is substituted or unsubstituted phenyl; in another embodiment R ⁵ is (C ₆ )aryl(C ₁ -C ₆ )alkyl , in other embodiments, R ⁵ is substituted or unsubstituted benzyl, in other embodiments, R ⁵ is benzyl, 2-fluorobenzyl, 3-fluorobenzyl, 4-fluorobenzyl, 2-chlorobenzyl , 3-chlorobenzyl, 4-chlorobenzyl, 3-phenylpropyl and phenethyl, preferably benzyl, 3-chlorobenzyl, 4-chlorobenzyl and 3-phenylpropyl. In some embodiments, R ⁵ is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, in other embodiments 5-methylfuran-2-ylmethyl, thiophene-2- ylmethyl and 2-(1H-indol-3-yl)ethyl, preferably selected from the group consisting of 5-methylfuran-2-ylmethyl and 2-(1H-indol-3-yl)ethyl. In some embodiments, R ⁵ is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl, in other embodiments furan-2-yl-methylthiomethyl, thiophene-2 -yl-methylthiomethyl, furan-2-yl-methoxymethyl, thiophen-2-yl-methoxymethyl, preferably furan-2-yl-methylthiomethyl. In some embodiments, R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, preferably (C ₆ )aryl(C ₁ -C ₆ )heteroalkyl; Preferably wherein aryl is selected from the group consisting of substituted or unsubstituted naphthyl and substituted or unsubstituted phenyl, more preferably wherein aryl is substituted or unsubstituted phenyl. Arylalkyl, heteroarylalkyl, heteroarylheteroalkyl and arylheteroalkyl are selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably halogen , more preferably chlorine or fluorine, even more preferably chlorine, even more preferably chlorine, preferably methyl, may be substituted by one or more identical or different substituents. Preferably the number of substituents in a substituted R5 group is 1, 2, ³ or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents in substituted R5 groups can be present ^on any position of the carbon atom, as generally described above for substituted alkyl groups.

In some embodiments, R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl, and in other embodiments (C ₁ -C ₂ )alkyl or (C ₁ - C ₂ )haloalkyl, in another embodiment (C1-C4)alkyl, preferably methyl, in another embodiment (C ₁ -C ₄ )haloalkyl, preferably CF ₃ . Alkyl may be optionally substituted by one or more same or different substituents selected from the group consisting of halogen and cyano, preferably halogen. Preferably the number of substituents in a substituted R6 group is ¹ , 2, 3 or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents in substituted R6 groups can be present ^on the carbon atom at any position, as generally described above for substituted alkyl groups.

In some embodiments, Y ¹ is selected from the group consisting of NB, CH-B and O, preferably NB and CH-B. In some embodiments Y ¹ is NB, in other embodiments Y ¹ is CH-B, in other embodiments Y ¹ is O.

In some embodiments, B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ - selected from the group consisting of C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C _{1 -C 6} )alkyl and N(R ⁷ )(R ⁸ ); ₁₀ ) from aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, In other embodiments, from (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, in other embodiments, ( (C ₆ )aryl, (C ₃ -C ₁₀ )heteroaryl and (C ₆ )aryl(C ₁ -C ₆ )alkyl. In some embodiments, B is (C ₆ -C ₁₀ )aryl, preferably wherein aryl is selected from the group consisting of substituted or unsubstituted naphthyl and substituted or unsubstituted phenyl, more preferably wherein aryl is substituted or unsubstituted phenyl. In some embodiments, B is phenyl, 3-methoxyphenyl, 4-fluorophenyl, 4-methoxyphenyl, 2-fluorophenyl, 2,5-dimethylphenyl, preferably phenyl, 4-fluorophenyl and 3- Methoxyphenyl, more preferably selected from the group consisting of phenyl and 3-methoxyphenyl. In some embodiments, B is (C ₃ -C ₁₀ )heteroaryl, in other embodiments B is 1-3 nitrogen atoms, preferably 1-2 nitrogen atoms, more preferably (C ₃ -C ₁₀ )heteroaryl containing 1 nitrogen atom, in another embodiment B consists of pyridin-2-yl, pyridin-3-yl, pyridin-4-yl is selected from the group, preferably pyridin-2-yl. In some embodiments, B is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, in other embodiments (C ₆ )aryl(C ₁ -C ₆ )alkyl , in other embodiments substituted or unsubstituted benzyl, in other embodiments benzyl, 2-chlorobenzyl, 2-fluorobenzyl, 3-chlorobenzyl, 3-fluorobenzyl, 4-chlorobenzyl, 4- fluorobenzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2,3-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl and 3,5-dimethoxybenzyl, preferably benzyl and It is selected from the group consisting of 3-chlorobenzyl, more preferably benzyl. In some embodiments, B is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably wherein the heteroaryl moiety is an oxygen-containing heteroaryl such as furanyl, or Sulfur-containing heteroaryls such as thiophene, or nitrogen-containing heteroaryls such as indole or diazaindole. In other embodiments, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl is pyridin-2-ylmethyl, pyridin-3-ylmethyl, pyridin-4-ylmethyl, 5-methylfuran-2- ylmethyl, furan-2-ylmethyl, furan-3-ylmethyl, thiophen-2-ylmethyl and thiophen-3-ylmethyl, preferably pyridin-2-ylmethyl and furan-2-ylmethyl, more preferably pyridin-2- is ylmethyl. In some embodiments, B is (C ₁ -C ₆ )alkyl, preferably (C ₁ -C ₄ )alkyl, in other embodiments methyl, ethyl, propyl, isopropyl, butyl, sec- It is selected from the group consisting of butyl and tert-butyl, preferably methyl or ethyl. In some embodiments, B is N(R ⁷ )(R ⁸ ), preferably wherein R ⁷ is hydrogen and R ⁸ is defined below, preferably wherein R ⁷ is (C ₁ -C ₆ )alkyl, preferably methyl, and R ⁸ is defined below. Aryl, heteroaryl, arylalkyl, heteroarylalkyl, alkyl and N(R ⁷ )(R ⁸ ) groups are halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , It may be substituted by one or more identical or different substituents, preferably selected from the group consisting of halogen and ^-OR6 . Preferably the number of substituents in a substituted B group is 1, 2, 3 or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents on substituted B groups are carbon at any position as generally described above for substituted aryl, heteroaryl, arylalkyl, heteroarylalkyl, alkyl and N(R ⁷ )(R ⁸ ) groups. Can exist on atoms.

B ¹ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl or (C ₃ -C ₁₀ )heteroaryl(C ₁ -C6)alkyl. In some embodiments, B ¹ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, and in other embodiments (C ₆ )aryl(C ₁ -C ₆ )alkyl. in another embodiment (C ₁₀ )aryl(C ₁ -C ₆ )alkyl, in another embodiment substituted or unsubstituted benzyl, in another embodiment substituted or unsubstituted naphthalene- 2-ylmethyl, in other embodiments naphthalen-2-ylmethyl, benzyl, 2-chlorobenzyl, 2-fluorobenzyl, 3-chlorobenzyl, 3-fluorobenzyl, 4-chlorobenzyl, 4-fluorobenzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2,3-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, naphthalen-2-ylmethyl and 3,5-dimethoxybenzyl, preferably is selected from the group consisting of benzyl, 4-fluorobenzyl, 2,4-dimethoxybenzyl and 3-methoxybenzyl, more preferably benzyl. In some embodiments, B ¹ is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably wherein the heteroaryl moiety is an oxygen-containing heteroaryl such as furanyl, or a sulfur-containing heteroaryl such as thiophene, or a nitrogen-containing heteroaryl such as pyridine, indole or diazaindole. In other embodiments, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl is pyridin-2-ylmethyl, pyridin-3-ylmethyl, pyridin-4-ylmethyl, 5-methylfuran-2- ylmethyl, furan-2-ylmethyl, furan-3-ylmethyl, thiophen-2-ylmethyl and thiophen-3-ylmethyl, preferably pyridin-2-ylmethyl and furan-2-ylmethyl, more preferably pyridin-2- is ylmethyl. _{1 _ _ _} ^{_ _} It may be substituted with one or more same or different substituents. Preferably the number of substituents in the substituted B ¹ group is 1, 2, 3 or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents in substituted B ¹ groups can be present on any position of the carbon atom, as described above generally for substituted arylalkyl and heteroarylalkyl groups.

In some embodiments, B ² is selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably halogen and —OR ⁶ , (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl substituted by one or more same or different substituents. In some embodiments, B ² is (C ₆ )aryl(C ₁ -C ₆ )alkyl, in other embodiments (C ₁₀ )aryl(C ₁ -C ₆ )alkyl, in others in embodiments substituted or unsubstituted benzyl; in other embodiments substituted or unsubstituted naphthalen-2-ylmethyl; in other embodiments 2-chlorobenzyl, 2-fluorobenzyl, 3-chloro benzyl, 3-fluorobenzyl, 4-chlorobenzyl, 4-fluorobenzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2,3-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4- It is selected from the group consisting of dimethoxybenzyl and 3,5-dimethoxybenzyl, preferably 4-fluorobenzyl and 3-methoxybenzyl. In some embodiments, B ² is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably wherein the heteroaryl moiety is an oxygen-containing heteroaryl such as furanyl, or a sulfur-containing heteroaryl such as thiophene, or a nitrogen-containing heteroaryl such as pyridine, indole or diazaindole. In other embodiments, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl is pyridin-2-ylmethyl, pyridin-3-ylmethyl, pyridin-4-ylmethyl, 5-methylfuran-2- ylmethyl, furan-2-ylmethyl, furan-3-ylmethyl, thiophen-2-ylmethyl and thiophen-3-ylmethyl, preferably pyridin-2-ylmethyl and furan-2-ylmethyl, more preferably pyridin-2- is ylmethyl. Heteroarylalkyl groups are one or more identical groups selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably halogen and —OR ⁶ . or may be substituted by different substituents. Preferably the number of substituents in the substituted B2 group is 1, ² , 3 or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents in substituted ^B2 groups can be present at any position on the carbon atom, as generally described above for substituted arylalkyl and heteroarylalkyl groups.

B ³ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C6)alkyl, (C1-C6)alkyl and N(R ⁷ ) (R ⁸ ). In some embodiments, B ³ is selected from the group consisting of (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C6)alkyl In other embodiments, B ³ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl. In some embodiments, B ³ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, in other embodiments (C ₆ )aryl(C ₁ -C ₆ )alkyl, In another embodiment substituted or unsubstituted benzyl, in another embodiment substituted or unsubstituted benzyl, in another embodiment substituted or unsubstituted naphthalen-2-ylmethyl, in another embodiment benzyl, naphthalene- 2-ylmethyl, 2-chlorobenzyl, 2-fluorobenzyl, 3-chlorobenzyl, 3-fluorobenzyl, 4-chlorobenzyl, 4-fluorobenzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2 ,3-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl and 3,5-dimethoxybenzyl, preferably benzyl and 3-chlorobenzyl, more preferably benzyl. In some embodiments, B ³ is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably wherein the heteroaryl moiety is an oxygen-containing heteroaryl such as furanyl, or a sulfur-containing heteroaryl such as thiophene, or a nitrogen-containing heteroaryl such as pyridine, indole or diazaindole. In other embodiments, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl is pyridin-2-ylmethyl, pyridin-3-ylmethyl, pyridin-4-ylmethyl, 5-methylfuran-2- ylmethyl, furan-2-ylmethyl, furan-3-ylmethyl, thiophen-2-ylmethyl and thiophen-3-ylmethyl, preferably pyridin-2-ylmethyl and furan-2-ylmethyl, more preferably pyridin-2- is ylmethyl. In some embodiments, B ³ is (C ₁ -C ₆ )alkyl, preferably (C ₁ -C ₄ )alkyl, in other embodiments methyl, ethyl, propyl, isopropyl, butyl, sec -butyl and tert-butyl, preferably methyl or ethyl. In some embodiments, B ³ is N(R ⁷ )(R ⁸ ), preferably wherein R ⁷ is hydrogen and R ⁸ is defined below, preferably wherein R ⁷ is (C ₁ -C ₆ )alkyl, preferably methyl, and R ⁸ is defined below. Arylalkyl, heteroarylalkyl, alkyl and N(R ⁷ )(R ⁸ ) groups are halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably halogen and — It may be substituted by one or more same or different substituents selected from the group consisting of ^OR6 . Preferably the number of substituents in the substituted B3 group is 1, 2, ³ or 4, more preferably 1, 2 or 3, such as 1 or 2. Substituents in substituted ^B3 groups may be on any position of the carbon atom, as generally described above for substituted arylalkyl, heteroarylalkyl, alkyl and N( ^R7 ) ⁽ R8) groups. obtain.

In some embodiments, R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl, in other embodiments hydrogen or (C ₁ -C ₄ )alkyl, in other embodiments hydrogen or (C ₃ -C ₄ )alkyl, in other embodiments hydrogen or (C ₁ -C ₂ )alkyl, in other embodiments R ⁷ is hydrogen, in other embodiments R ⁷ is (C ₁ -C ₆ )alkyl, preferably (C ₁ -C ₄ )alkyl, more preferably (C ₁ -C ₂ )alkyl, even more preferably methyl. Alkyl may be optionally substituted by one or more same or different substituents selected from the group consisting of halogen and cyano, preferably halogen. Preferably the number of substituents in a substituted ^R7 group is 1, 2, 3 or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents in substituted ^R7 groups can be present on the carbon atom at any position, as generally described above for substituted alkyl groups.

In some embodiments, R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ —C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl; in other embodiments, (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ ) from heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, in other embodiments, (C ₆ —C ₁₀ )aryl and (C ₃ -C ₁₀ )heteroaryl, in other embodiments, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl ( C ₁ -C ₆ )alkyl, in other embodiments, (C ₆ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C 6 )alkyl ₁₀ ) selected from heteroaryl(C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl; In some embodiments, R ⁸ is (C ₆ -C ₁₀ )aryl, preferably wherein aryl is selected from the group of substituted or unsubstituted naphthyl and substituted or unsubstituted phenyl, more preferably wherein aryl is substituted or unsubstituted phenyl. In some embodiments, R ⁸ is (C ₃ -C ₁₀ )heteroaryl, in other embodiments (C ₃ -C ₁₀ )heteroaryl is 1-3 nitrogen atoms, preferably It contains 1 to 2 nitrogen atoms, more preferably 1 nitrogen atom, eg pyridin-2-yl, pyridin-3-yl, pyridin-4-yl. In some embodiments, R ⁸ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, and in other embodiments (C ₆ )aryl(C ₁ -C ₆ )alkyl. , in other embodiments substituted or unsubstituted benzyl, in other embodiments benzyl, 2-chlorobenzyl, 2-fluorobenzyl, 3-chlorobenzyl, 3-fluorobenzyl, 4-chlorobenzyl, 4 -fluorobenzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2,3-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl and 3,5-dimethoxybenzyl, preferably benzyl and 3-chlorobenzyl, more preferably benzyl. In some embodiments, R ⁸ is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably wherein the heteroaryl moiety is an oxygen-containing heteroaryl such as furanyl, or sulfur containing heteroaryl such as thiophene or nitrogen containing heteroaryl such as indole or diazaindole. In other embodiments, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl is pyridin-2-ylmethyl, pyridin-3-ylmethyl, pyridin-4-ylmethyl, 5-methylfuran-2- ylmethyl, furan-2-ylmethyl, furan-3-ylmethyl, thiophen-2-ylmethyl and thiophen-3-ylmethyl, preferably pyridin-2-ylmethyl and furan-2-ylmethyl, more preferably pyridin-2- is ylmethyl. In some embodiments, R ⁸ is (C ₁ -C ₆ )alkyl, preferably (C ₁ -C ₄ )alkyl, in other embodiments methyl, ethyl, propyl, isopropyl, butyl, sec -butyl and tert-butyl, preferably selected from the group consisting of methyl or ethyl. Aryl, heteroaryl, arylalkyl, heteroarylalkyl and alkyl groups are the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably halogen and —OR ⁶ It may be substituted with one or more of the same or different substituents selected from. Preferably the number of substituents in a substituted B group is 1, 2, 3 or 4, more preferably 1, 2 or 3, eg 1 or 2. Substituents on substituted B groups can be present on any carbon atom, as generally described above for substituted aryl, heteroaryl, arylalkyl, heteroarylalkyl and alkyl groups.

In some embodiments, p is an integer between 1 and 2, in other embodiments p is 1, and in other embodiments p is 2.

In some embodiments q is an integer between 1 and 3, in other embodiments q is 1, 2 or 3, in other embodiments q is an integer between 1 and 2 and in other embodiments q is 1, in other embodiments q is 2, and in other embodiments q is 3.

In some embodiments p is 1 and q is 1, in other embodiments p is 1 and q is 2, in other embodiments p is 1 and q is 3, in other embodiments p is 2 and q is 2, in other embodiments p is 2 and q is 3.

（式中、Ｒ^１、Ｒ^２、Ｒ^３、Ｘ、Ｙ、Ｚ、Ｂ、ｎ、ｐ及びｑは、上記及び下記のように定義される）
による構造を有する化合物である。
Ｙは、ＣＨ又はＮである。いくつかの実施形態において、ＹはＣＨであり、他の実施形態において、ＹはＮである。 In some embodiments, the compound having a structure according to formula (I) has formula (II):

(wherein R ¹ , R ² , R ³ , X, Y, Z, B, n, p and q are defined above and below)
is a compound having a structure according to
Y is CH or N; In some embodiments Y is CH, and in other embodiments Y is N.

（式中、Ｙ、Ｂ、ｎ及びｑは、上記及び下記のように定義される）
による構造を有する化合物である。 In some embodiments, the compound having a structure according to formula (I) has formula (III):

(wherein Y, B, n and q are defined above and below)
is a compound having a structure according to

（式中、Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４、Ｒ^５、Ｘ、Ｚ及びｎは、上記及び下記のように定義される）
による構造を有する化合物である。 In some embodiments, the compound having a structure according to formula (I) has formula (IV):

(wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , X, Z and n are defined above and below)
is a compound having a structure according to

の基であり、ｐは１であり、ｑは１であり、他の実施形態において、Ａは、下記式：

の基であり、ｐは２であり、ｑは１であり、他の実施形態において、Ａは、下記式：

の基であり、ｐは２であり、ｑは２であり、他の実施形態において、Ａは、下記式：

の基であり、ｐは２であり、ｑは３である。 In some embodiments, A is of the formula:

p is 1 and q is 1, and in other embodiments A is the group

p is 2 and q is 1, and in another embodiment A is the group

p is 2 and q is 2, and in other embodiments A is the group

, p is 2 and q is 3.

の基からなる群より選択され；
Ｂ^１が、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル又は（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ６）アルキルであり、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｂ^２が、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換される（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキルであるか、又は、
Ｂ^２が、場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ^６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換される（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキルであり；
Ｂ^３が、水素、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_１－Ｃ_６）アルキル及びＮ（Ｒ^７）（Ｒ^８）からなる群より選択され、これらは場合により、ハロゲン、（Ｃ_１－Ｃ_４）アルキル、（Ｃ_１－Ｃ_４）ハロアルキル及び－ＯＲ_６からなる群より選択される、１つ以上の同一又は異なる置換基によって置換され；
Ｒ^６が、（Ｃ_１－Ｃ_４）アルキル又は（Ｃ_１－Ｃ_４）ハロアルキルであり；
Ｒ^７が、水素又は（Ｃ_１－Ｃ_６）アルキルであり；
Ｒ^８が、（Ｃ_６－Ｃ_１０）アリール、（Ｃ_３－Ｃ_１０）ヘテロアリール、（Ｃ_６－Ｃ_１０）アリール（Ｃ_１－Ｃ_６）アルキル、（Ｃ_３－Ｃ_１０）ヘテロアリール（Ｃ_１－Ｃ_６）アルキル及び（Ｃ_１－Ｃ_６）アルキルからなる群より選択され；
ｐが１であり、かつ
ｑが、１と２の間の整数であるか、又は、
ｐが２であり、かつ
ｑが３である
ことを特徴とする化合物、又はその薬学的に許容される塩、溶媒和物若しくは水和物である。 In some embodiments, compounds having a structure according to formula (I) are
X is selected from the group consisting of N and CH;
R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) selected from the group consisting of aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl;
R ² and R ³ are independently selected from the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl;
n is an integer between 0 and 3;
Z is selected from the group consisting of C=O, C=S and _CH2 ;
A is the following formula:

selected from the group consisting of
B ¹ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl or (C ₃ -C ₁₀ )heteroaryl(C ₁ -C6)alkyl, which are optionally halogen, (C ₁ substituted by one or more identical or different substituents selected from the group consisting of —C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ ;
_B ^{2 is} _{substituted ( C 6} -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, or
B ² is optionally substituted by one or more same or different substituents selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ is (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl;
B ³ is hydrogen, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl and N(R ⁷ )(R ⁸ ), which are optionally selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ₆ , substituted by one or more identical or different substituents;
R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl;
R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl( selected from the group consisting of C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl;
p is 1 and q is an integer between 1 and 2, or
A compound, or a pharmaceutically acceptable salt, solvate or hydrate thereof, characterized in that p is 2 and q is 3.

In some embodiments, the compound having the structure according to formula (I) is a compound having the structure according to formula (II)
X is selected from the group consisting of N and CH, preferably N, preferably CH;
R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably hydrogen, (C ₁ -C ₆ )alkyl , (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl;
R ² and R ³ are independently hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl, preferably (C ₁ -C ₆ ) selected from the group consisting of alkyl and (C ₁ -C ₆ )haloalkyl;
n is an integer between 0 and 3, preferably 1-3, more preferably 1-2, even more preferably 2;
Z is selected from the group consisting of C=O, C=S and _CH2 , preferably C=O;
Y is N or CH;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl and N(R ⁷ )(R ⁸ ) which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably substituted by one or more identical or different substituents selected from the group consisting of halogen and —OR ⁶ ;
R ⁷ is hydrogen or (C ₁ -C ₆ )alkyl, preferably hydrogen;
R ⁸ is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl( C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl, preferably (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ ) alkyl, more preferably selected from the group consisting of (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl;
p is an integer between 1 and 2;
A compound wherein q is an integer between 1 and 3, or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In some embodiments, compounds having a structure according to Formula (I) or Formula (II) are
R ¹ is hydrogen, (C ₁ -C ₄ )alkyl, (C ₃ -C ₆ )cycloalkyl, (C ₆ -C ₁₀ )aryl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl selected from the group consisting of;
R ² and R ³ are independently selected from the group consisting of (C ₁ -C ₄ )alkyl and (C ₁ -C ₄ )haloalkyl;
n is an integer between 1 and 3, preferably between 1 and 2, more preferably 2;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl, which are optionally selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably halogen It is characterized by being substituted by one or more selected identical or different substituents.

In some embodiments, the compound having the structure according to formula (I) is a compound having the structure according to formula (III)
n is an integer between 1 and 3, preferably between 1 and 2, more preferably 2;
Y is N or CH;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , preferably substituted by one or more identical or different substituents selected from the group consisting of halogen and —OR ⁶ ;
A compound wherein q is an integer between 1 and 3, preferably between 1 and 2, more preferably 2, or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In some embodiments, the compound having the structure according to formula (I) is a compound having the structure according to formula (II)
X is selected from the group consisting of N and CH;
R ¹ is hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, (C ₆ -C ₁₀ )aryl, (C ₆ -C ₁₀ ) aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl and (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, preferably hydrogen and (C ₁ -C ₆ )alkyl , more preferably selected from the group consisting of (C ₁ -C ₆ )alkyl;
R ² and R ³ are independently hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl and (C ₃ -C ₈ )cycloalkyl, preferably (C ₁ -C ₆ ) selected from the group consisting of alkyl and (C ₁ -C ₆ )haloalkyl, more preferably (C ₁ -C ₆ )alkyl;
n is an integer between 0 and 3, preferably 1-3, more preferably 1-2, even more preferably 2;
Z is selected from the group consisting of C=O, C=S and _CH2 , preferably C=O;
R ⁴ is hydrogen or (C ₁ -C ₆ )alkyl, preferably hydrogen;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ , optionally substituted by one or more identical or different substituents selected from the group consisting of halogen;
A compound wherein R ⁶ is (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )haloalkyl, or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In some embodiments, compounds of formula (I) are:
1-(4-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperazin-1-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperazin-1-yl)propan-1-one,
(R)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
(S)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-cyclopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5- dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-phenyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(1-(3-benzyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( 4-benzylpiperidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(4-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)ethan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propane-1-thione,
6-(4-(3-(4-benzylpiperidin-1-yl)propyl)-3,5-dimethyl-1H-pyrazol-1-yl)-3-methyl-[1,2,4]triazolo[4 , 3-b]pyridazine,
1-(azepan-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H- pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-morpholino propane-1-one,
N-(3-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)-N-methylpropanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(furan-2-ylmethyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-phenylpropan-2-yl)piperazin-1-yl)propan-1-one,
1-(4-(3,4-dimethoxybenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b ] pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(naphthalen-2-ylmethyl)piperazin-1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylazetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl )-1H-pyrazol-4-yl)propan-1-one,
1-(3-(benzylamino)azetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6 -yl)-1H-pyrazol-4-yl)propan-1-one,
N-benzyl-3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole-4-carboxamide,
(3-benzylpyrrolidin-1-yl)(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole- 4-yl)methanone,
1-(3-benzylpyrrolidin-1-yl)-3-(5-isopropyl-3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6- yl)-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-5 -(trifluoromethyl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperazin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorophenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(4 -fluorophenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( pyrrolidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(3 - methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(4 - methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-phenylpiperazine -1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(pyridine -2-yl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( (5-methylfuran-2-yl)methyl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-fluorophenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 3-phenylpropyl)propanamide,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-benzylpiperidine -1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2,5-dimethylphenyl)piperazin-1-yl)propan-1-one,
N-(4-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( thiophen-2-ylmethyl)propanamide,
N-(2-(1H-indol-3-yl)ethyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propanamide,
N-benzyl-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) propanamide,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-N-(3-phenylpropyl ) propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-ethylpiperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-methylpiperidine -1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(pyrrolidin-1- yl) propane-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 2-fluorobenzyl)propanamide, and
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 2-((furan-2-ylmethyl)thio)ethyl)propanamide,
is selected from a list consisting of

In some embodiments, compounds of formula (I) are:
1-(4-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperazin-1-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperazin-1-yl)propan-1-one,
(R)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
(S)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-cyclopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5- dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-phenyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(1-(3-benzyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( 4-benzylpiperidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(4-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)ethan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propane-1-thione,
6-(4-(3-(4-benzylpiperidin-1-yl)propyl)-3,5-dimethyl-1H-pyrazol-1-yl)-3-methyl-[1,2,4]triazolo[4 , 3-b]pyridazine,
1-(azepan-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H- pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-morpholino propane-1-one,
N-(3-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)-N-methylpropanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(furan-2-ylmethyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-phenylpropan-2-yl)piperazin-1-yl)propan-1-one,
1-(4-(3,4-dimethoxybenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b ] pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(naphthalen-2-ylmethyl)piperazin-1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylazetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl )-1H-pyrazol-4-yl)propan-1-one,
1-(3-(benzylamino)azetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6 -yl)-1H-pyrazol-4-yl)propan-1-one,
N-benzyl-3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole-4-carboxamide,
(3-benzylpyrrolidin-1-yl)(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole- 4-yl)methanone,
1-(3-benzylpyrrolidin-1-yl)-3-(5-isopropyl-3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6- yl)-1H-pyrazol-4-yl)propan-1-one, and
1-(3-benzylpyrrolidin-1-yl)-3-(3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-5 -(trifluoromethyl)-1H-pyrazol-4-yl)propan-1-one,
is selected from a list consisting of

In some embodiments, compounds of formula (I) are:
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperazin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorophenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(4 -fluorophenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( pyrrolidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(3 - methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(4 - methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-phenylpiperazine -1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(pyridine -2-yl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( (5-methylfuran-2-yl)methyl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-fluorophenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 3-phenylpropyl)propanamide,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-benzylpiperidine -1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2,5-dimethylphenyl)piperazin-1-yl)propan-1-one,
N-(4-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( thiophen-2-ylmethyl)propanamide,
N-(2-(1H-indol-3-yl)ethyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propanamide,
N-benzyl-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) propanamide,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-N-(3-phenylpropyl ) propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-ethylpiperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-methylpiperidine -1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(pyrrolidin-1- yl) propane-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 2-fluorobenzyl)propanamide, and
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 2-((furan-2-ylmethyl)thio)ethyl)propanamide,
is selected from a list consisting of

In some embodiments, compounds of formula (I) are:
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperazin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorophenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(4 -fluorophenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( pyrrolidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(3 - methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(4 - methoxyphenyl)piperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-phenylpiperazine -1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-(pyridine -2-yl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( (5-methylfuran-2-yl)methyl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-fluorophenyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 3-phenylpropyl)propanamide,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-benzylpiperidine -1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2,5-dimethylphenyl)piperazin-1-yl)propan-1-one,
N-(4-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( thiophen-2-ylmethyl)propanamide,
N-(2-(1H-indol-3-yl)ethyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propanamide,
N-benzyl-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) propanamide,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-N-(3-phenylpropyl ) propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-ethylpiperazin-1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(4-methylpiperidine -1-yl)propan-1-one,
3-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-(pyrrolidin-1- yl) propane-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 2-fluorobenzyl)propanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-N-( 2-((furan-2-ylmethyl)thio)ethyl)propanamide,
1-(4-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperazin-1-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenylpiperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-yl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxybenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(4-fluorobenzyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperidin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(pyridin-2-ylmethyl)piperazin-1-yl)propan-1-one,
(R)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
(S)-1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- 6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-cyclopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5- dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-phenyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(1-(3-benzyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( 4-benzylpiperidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(4-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)ethan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propane-1-thione,
6-(4-(3-(4-benzylpiperidin-1-yl)propyl)-3,5-dimethyl-1H-pyrazol-1-yl)-3-methyl-[1,2,4]triazolo[4 , 3-b]pyridazine,
1-(azepan-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H- pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-morpholino propane-1-one,
N-(3-chlorobenzyl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole -4-yl)-N-methylpropanamide,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(furan-2-ylmethyl)piperazin-1-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(2-phenylpropan-2-yl)piperazin-1-yl)propan-1-one,
1-(4-(3,4-dimethoxybenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b ] pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(naphthalen-2-ylmethyl)piperazin-1-yl)propan-1-one,
1-(4-(3-chlorobenzyl)piperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylazetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl )-1H-pyrazol-4-yl)propan-1-one,
1-(3-(benzylamino)azetidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6 -yl)-1H-pyrazol-4-yl)propan-1-one,
N-benzyl-3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole-4-carboxamide,
(3-benzylpyrrolidin-1-yl)(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole- 4-yl)methanone,
1-(3-benzylpyrrolidin-1-yl)-3-(5-isopropyl-3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6- yl)-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-5 -(trifluoromethyl)-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-(3-methoxyphenyl)piperazin-1-yl)propan-1-one,
1-benzyl-4-(3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazole-4 -yl)propanoyl)piperazin-2-one,
1-(4-benzylpiperidin-1-yl)-3-(3,5-diethyl-1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl -1H-pyrazol-4-yl)propan-1-one,
1-(2,2-dimethylpyrrolidin-1-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5 -dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(3,3-dimethylpyrrolidin-1-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5 -dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-diethyl-1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-diethyl-1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 3,3-dimethylpyrrolidin-1-yl)propan-1-one,
1-((1S)-2-azabicyclo[2.2.1]heptan-2-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)propan-1-one,
(1S,5R)-3-(3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazole -4-yl)propanoyl)-2,3,4,5-tetrahydro-1,5-methanopyrido[1,2-d][1,4]diazepin-7(1H)-one,
1-((1S)-2-azabicyclo[2.2.1]heptan-2-yl)-3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-(trifluoromethyl)-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) -1-(pyrrolidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl -1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl -1H-pyrazol-4-yl)propan-1-one,
3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 3,3-dimethylpyrrolidin-1-yl)propan-1-one,
3-(1-(3-ethyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( pyrrolidin-1-yl)propan-1-one,
3-(1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( pyrrolidin-1-yl)propan-1-one,
3-(3,5-diethyl-1-(3-isopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 3,3-dimethylpyrrolidin-1-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-diethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(4-benzyl-1,4-diazepan-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine -6-yl)-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(3-isopropyl-5-methyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine-6- yl)-1H-pyrazol-4-yl)propan-1-one,
3-(1-(3-benzyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5-dimethyl-1H-pyrazol-4-yl)-1-( 4-benzylpiperidin-1-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-3-(1-(3-cyclopropyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-3,5- dimethyl-1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperazin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(4-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(3-benzylpiperazin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpiperidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
1-(3-benzylpyrrolidin-1-yl)-2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl) -1H-pyrazol-4-yl)propan-1-one,
1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl ) propane-1-one,
2-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-( 4-phenethylpiperidin-1-yl)propan-1-one, and
3-(3,5-dimethyl-1-(3-methylimidazo[1,2-b]pyridazin-6-yl)-1H-pyrazol-4-yl)-1-(4-phenethylpiperidin-1-yl ) propane-1-one,
is selected from a list consisting of

A selection of compounds within the scope of the invention, or uses within the methods of the invention, are listed in Table 1 below.

In a further aspect, the invention provides a compound of the invention (especially those identified above with respect to any of formulas (I), (II), (III) and (IV)) for use as a medicament. , as well as the compounds of Table 1.

As is evident from the examples, the present inventors have found that the compounds of the invention inhibit or reduce the TRAF6-Ubc13 interaction, in particular that the compounds of the invention inhibit or reduce the enzymatic activity of TRAF6. rice field. In some embodiments, the compounds of the invention are selective inhibitors of TRAF6. In some embodiments, the compounds of the present invention have pharmacological properties (bioavailability, toxicity, side effects, dosage, patient compliance, suitability, stability, half-life, etc.).

In preferred compounds of the present invention, any one or more structural elements such as groups, substituents and numbers are defined in any of the preferred definitions of that element or in any particular embodiment and/or Elements mentioned as examples can have one or more specific meanings, where one or more preferred definitions and embodiments and/or all combinations of specific meanings are subject matter of the present invention. Also, for all preferred compounds of Formula I, Formula II, Formula III and Formula IV, all their stereoisomeric forms and mixtures of stereoisomeric forms in all ratios, and physiologically acceptable salts thereof , as well as physiologically acceptable solvates of any of them, are the subject of the present invention. Similarly, the various groups and numbers in the general definitions of all the specific compounds disclosed herein (e.g., the compounds of the Examples (compounds of Formula I, Formula II, Formula III, and Formula IV) are )), which represents an embodiment of the present invention, which has a particular meaning present in a particular compound of )), all stereoisomeric forms thereof and mixtures of stereoisomeric forms in all ratios, as well as their physiological and the physiologically acceptable solvates of any of them are subject matter of the present invention. In particular, the subject of the present invention is both the free compound form and all physiologically acceptable salt forms thereof, whether disclosed as the free compound and/or as a specific salt (and , where a specific salt is disclosed, in addition to this specific salt form and physiologically acceptable solvate forms thereof), all specific compounds disclosed herein .

The compounds of the invention can be prepared as described below, or by methods analogous thereto, which are readily apparent and accessible to those skilled in the art of organic synthesis. Synthetic access to compounds according to the invention, ie compounds of formulas (I), (II), (III) and (IV), is further illustrated by the exemplary compounds in the description.

In the course of synthesizing a compound of formula I, a heterocyclic compound of formula V can be treated with hydrazine to give the corresponding compound of formula VI and then condensed with a compound of formula VII to saponify to a compound of formula IX. (The groups X, R ¹ , R ² and R ³ and the number n are defined as in compounds of formula I). LG ₁ is any leaving group (halide, tosylate, etc.) suitable for chemical conversion of compound V to compound VI. R ^L is any residue suitable for the chemical reaction of compound VI and compound VII to give compound VIII, i.e., stable under the reaction conditions and cleavable under different reaction conditions, allowing the preparation of compound IX. residues such as alkyl groups such as methyl or ethyl. Additional functional groups may be present in protected form or in the form of precursor groups which are later converted to the final group. The hydrazine applied for the conversion of compound V to compound VI can be hydrazine or a hydrazine equivalent such as hydrazine hydrate or hydrazine salt. The reaction of hydrazine or a hydrazine equivalent with compound V is usually carried out at a temperature of about 0°C to 150°C, preferably 30°C to 100°C. Hydrolytic work-up of the reaction mixture, or isolation of the product by filtration, as with all reaction work-up in the preparation of compounds of formula I, can generally be carried out under standard conditions, followed by A compound of VI is obtained.

The condensation reaction of compound VI and compound VII is preferably carried out under acidic conditions, such as in acidic acid as solvent, to give compound of formula VIII. The reaction is usually carried out at a temperature of about 30°C to 180°C, preferably 50°C to 150°C.

Compounds of formula VIII are converted to compounds of formula IX under acidic or basic conditions, preferably under basic conditions. Saponification can be carried out under acidic conditions by treating a compound of formula VIII with a strong acid such as HBr in a suitable solvent. Saponification can be carried out under basic conditions by treating the compound of Formula VIII with a base such as NaOH, LiOH or KOH in a suitable solvent. The reaction is usually carried out at a temperature of about 0°C to 150°C, preferably 30°C to 130°C.

Compounds of formula IX are then coupled with compounds of formula X under standard conditions for amide bond formation to provide compounds of formula XI. The groups R ¹ , R ² , R ³ , X and Y and the numbers n, p and q are defined analogously as in the compounds of the formula I, further functional groups being added in protected form or later finally may be present in the form of precursor groups which are converted into the groups. Compounds of formula IX are as defined above. For amide bond formation, the carboxylic acid of Formula IX is typically converted to a reactive derivative, which can be isolated or prepared in situ, or activated in situ by conventional amide coupling reagents.

For example, compounds of formula IX can be converted to carboxylic acid chlorides, to reactive esters, or to ethyl chloroformate or isobutyl chloroformate by treatment with thionyl chloride, oxalyl chloride or (1-chloro-2-methylpropenyl)-dimethylamine. can be converted to the mixed anhydride by treatment with an alkyl chloroformate such as Alternatively, the compound of formula IX can be prepared as propanephosphonic anhydride, N,N'-carbonyldiazole such as N,N'-carbonyldiimidazole (CDI)), N,N'-diisopropylcarbodiimide (DIC), N , N′-dicyclohexylcarbodiimide (DCC) or N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), 1-hydroxybenzotriazole (HOBT) or 1-hydroxy-7- carbodiimides in combination with additives such as azabenzotriazole (HOAT); O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), O-( 7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-(benzotriazol-1-yl)-N,N,N', Uronium such as N'-tetramethyluronium hexafluorophosphate (HBTU) or O-(cyano(ethoxycarbonyl)methyleneamino)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TOTU) or (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP), or bromotri It can be activated with reagents such as phosphonium-based coupling reagents such as bromothpyrrolidinophosphonium hexafluorophosphate (PyBroP). Activation of compounds of formula IX and reaction of activated compounds of formula IX, i.e. reactive carboxylic acid derivatives, with compounds of formula X are generally carried out in an inert solvent such as tetrahydrofuran (THF), dioxane or 1, 2 ethers such as dimethoxyethane (DME), or hydrocarbons such as toluene, chlorinated hydrocarbons such as dichloromethane or chloroform, or dimethylformamide (DMF) or N-methylpyrrolidin-2-one (NMP) amides, etc., or in a mixture of solvents, a suitable base, for example a tertiary amine such as triethylamine, ethyl-diisopropylamine, N-methylmorpholine or pyridine, or a basic such as sodium carbonate, potassium carbonate or cesium carbonate. It is carried out at a temperature of about 0° C. to about 60° C. in the presence of an alkali metal compound or the like. Carboxylic acids of formula IX, or compounds containing a carboxylic acid derivative group, such as a carboxylic acid chloride group, in place of the carboxylic acid group shown in formula IX can be obtained from the corresponding esters (for example, 5-ethyl- by saponification of 4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid).

When all groups of formula XI have the desired meanings included in the definition of compounds of formula I, the compounds of formula XI thus obtained are already final compounds of formula I. Where any group of formula XI is present in protected form or in the form of a precursor group, the compound of formula XI thus obtained may be, by removal of the protecting group and/or transformation of any other group, finally It can be converted into the desired Formula I compound. As mentioned above, in any one or more steps in the synthesis of compounds of formula I, functional groups may be in protected form or in precursor groups to avoid undesired course of reactions, i.e. side reactions. can exist in the form Protecting groups can be removed and precursor groups can be transformed at other stages of the synthesis as well as at the final stage of the synthesis of compounds of formula I. Suitable protecting groups and respective synthetic strategies and details regarding their introduction and removal are well known to those skilled in the art, see, for example, P. G. M. Wuts and T. W. Greene, Greene's Protective Groups in Organic Synthesis, 4. ed. (2007), John Wiley Found in & Sons.

In addition, various other transformations of functional groups can be carried out in compounds of formula I, or compounds of formula XI, or other compounds occurring in the synthesis of compounds of formula I, in order to obtain further compounds of formula I. .

Compounds of formula XI can also be converted to compounds of formula XII or XIII. The groups R ¹ , R ² , R ³ , X and Y and the numbers n, p and q are defined analogously as in the compounds of formula I, further functional groups being in protected form or later finalized may be present in the form of precursor groups which are converted into the groups. Compounds of formula XI are as defined above.

Conversion of the carbonyl group of the compound of formula XI to the thiocarbonyl group of the compound of formula XII can be accomplished using various reagents known in the art such as Lawesson's reagent, ammonium phosphorodithionate, or PSCl ₃ /H ₂ O/ It can be done with the Et ₃ N system. The solvent and reaction time applied will depend on the reagent or combination of reagents utilized.

Reduction of the carbonyl group of a compound of formula XI to a compound of formula XII can be achieved by, for example, platinum-catalyzed reduction, metal-mediated reduction such as zinc-catalyzed reduction, tris(pentafluorophenyl)borane B _{(C6F5)3 or chlorobis} (cyclooctene)iridium dimer [Ir(COE)2Cl] ₂ , or reduction by hydride transfer using reagents such as lithium aluminum hydride _{( LiAlH4} ). Various reagents known in the art can be used.

When all groups of formulas XII and XIII have the desired meanings included in the definition of compounds of formula I, the compounds of formulas XII and XIII thus obtained are already final compounds of formula I. Where any group of formulas XII and XIII is present in protected form or in the form of a precursor group, the compounds of formulas XII and XIII thus obtained may be subjected to removal of protecting groups and/or transformation of any other groups. can finally be converted to the desired compound of formula I by

Starting compounds and building blocks for the synthesis of compounds of formula I are either commercially available or may be prepared according to or analogous to procedures described in the literature.

The order in which groups are introduced during the synthesis of compounds of Formula I may differ from that outlined above. For example, a thiocarbonyl group of a compound of formula XII can be introduced by converting the carbonyl group of a compound of formula VIII to the corresponding thioester. In particular, other synthetic routes are indeed applicable to certain embodiments of the compounds disclosed herein. Those skilled in the art are familiar with, for example, March's Advanced Organic Chemistry (Michael B. Smith & Jerry March, Wiley-Interscience, 2000), The Practice of Medicinal Chemistry (Camile G. Wermuth, Academia Press, 2003), and Protective Groups in Organic Synthesis ( See general textbooks such as Theosora W. Greene & Peter G.M. Wuts; John Wiley & Sons Inc, 1999).

Pharmaceutical Compositions In a further aspect, the invention provides pharmaceutical compositions comprising a compound identified above under the heading "Compound" and one or more pharmaceutically acceptable excipients.

Compounds described in the present invention (in particular those identified above, such as compounds of formulas (I), (II), (III) and/or (IV)) and compounds of Table 1 are preferably It is administered to a patient in need thereof via a pharmaceutical composition. In one embodiment, the pharmaceutical composition comprises a compound as described above (e.g., compounds having general formulas (I), (II), (III) and/or (IV), and compounds of Table 1, or hydrates thereof) , solvates, salts, complexes, racemic mixtures, diastereomers, enantiomers or tautomers, or isotopically enriched forms of any of the foregoing) and one or more pharmaceutically acceptable excipients. include.

A pharmaceutical composition can be administered to an individual by any route, eg, enterally or parenterally.

As used herein, the terms "enteral administration" and "enterally administered" mean that the administered drug is taken up by the stomach and/or intestines. Examples of enteral administration include oral and rectal administration. The terms "parenteral administration" and "administered parenterally" as used herein refer to modes of administration other than enteral administration (usually by injection or topical application), including intravenous, Intramuscular, intraarterial, intrathecal, intracapsular, intraosseous, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intraarticular, subcapsular, intracerebral, intracerebroventricular, arachnoid Although including sub-, intraspinal, epidural and intrasternal administration (e.g. by injection and/or infusion), and topical administration (e.g. by epithelial, inhalation, or mucosal (e.g. buccal, sublingual or vaginal)) , but not limited to.

The compounds used in the present invention are generally applied in "pharmaceutically acceptable amounts" and "pharmaceutically acceptable preparations." Such compositions may contain salts, buffering agents, preservatives, carriers, and optionally other therapeutic agents.

The term "excipient" as used herein includes, for example, carriers, binders, lubricants, thickeners, surfactants, preservatives, emulsifiers, buffering agents, flavoring agents, coloring agents, is intended to indicate any substance that is not an active ingredient in the pharmaceutical composition (e.g., a therapeutically inactive ingredient that exhibits no therapeutic effect at the amounts/concentrations used), such as agents or antioxidants. .

A composition according to the present invention may contain a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any physiologically compatible solvents, dispersion media, coatings, tonicity and absorption delaying agents and the like. A "pharmaceutically acceptable carrier" can be in the form of a solid, semi-solid, liquid, or combinations thereof. Preferably, the carrier is suitable for enteral (eg, oral) or parenteral administration (eg, intravenous, intramuscular, subcutaneous, spinal or epidermal administration (eg, by injection or infusion)). Depending on the route of administration, the active compound, i.e. the compound used in the present invention, either alone or in combination with one or more additional active compounds, may be susceptible to the effects of acids and other natural conditions that may inactivate the active compound. It may be coated with a material that protects the active compound from the

Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions used according to the invention include water (eg water for injection), ethanol, polyols (eg glycerol, propylene glycol, polyethylene glycol and the like), salts , carbohydrates, sugar alcohols, or aqueous solutions of amino acids (e.g., saline or aqueous amino acids), and suitable mixtures and/or buffered forms thereof, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., For example, ethyl oleate). Proper fluidity is maintained, for example, by using a coating material such as lecithin, maintaining the required particle size (in the case of dispersions), and by using surfactants.

Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The use of such media and agents for pharmaceutically active compounds is well known in the art. Unless any conventional media or agent is incompatible with the active compound, its use in the pharmaceutical compositions employed by the present invention is contemplated.

Additional active compounds are the compounds used in the present invention (in particular those identified above, such as compounds of (I), (II), (III) and/or (IV) and compounds of Table 1; and a compound of Table 1, or incorporated into this composition), either before or after. In one embodiment, the pharmaceutical compositions described herein are compounds described above (e.g., compounds having general formulas (I), (II), (III) and/or (IV), and compounds of Table 1) , or hydrates, solvates, salts, complexes, racemic mixtures, diastereomers, enantiomers, or tautomers thereof, or isotopically enriched forms of any of the foregoing), at least one additional active compound. , and one or more pharmaceutically acceptable excipients.

"Additional active compounds" (not compounds having formulas (I), (II), (III) and/or (IV) and compounds of Table 1 identified herein) are cancer and/or or any compound that can be used in the treatment of immune disorders. Additional active compounds may induce additive or synergistic therapeutic effects.

The pharmaceutical compositions described herein include at least one may contain, for example, 1, 2, 3, 4, 5, 6, 7 or 8 additional active compounds. According to the present teachings, at least additional active compounds, such as anti-cancer agents and/or agents for treating immune disorders, are compounds having the structures of formulas (I), (II), (III) and/or (IV), or, together with the compounds of Table 1 above, can be formulated into a single pharmaceutical composition. Alternatively, the pharmaceutical compositions can be structured as a kit of parts, wherein a compound having the structure of Formulas (I), (II), (III) and/or (IV) or a compound of Table 1 is , is provided in a first formulation, and at least one additional active compound, e.g., an anticancer agent and/or an agent for the treatment of an immune disorder, is provided in a second formulation, i.e., a second pharmaceutical composition. . The first and second pharmaceutical compositions can be combined prior to use. In other words, prior to administration of the pharmaceutical composition, formulations containing additional active compounds are added to compounds having the structure of formulas (I), (II), (III) and/or (IV), or It can be added to a first pharmaceutical composition containing one compound. Alternatively, the present teachings administer a compound having a structure of Formula (I), (II), (III) and/or (IV), or a compound of Table 1 above, formulated into a first pharmaceutical composition. and then administering at least one additional active compound formulated in a second pharmaceutical composition. Pharmaceutical compositions can be administered simultaneously or sequentially. For example, a first pharmaceutical composition can be administered at a first time point and a second pharmaceutical composition can be administered at a second time point, where the time points are, for example, 0, or 1 , up to 2, 3, 4, 5, or 10 minutes, up to 1, 2, 3, 4, 5, or 10 hours, up to 1, 2, 3, 4, 5, or 10 days, 1, 2, 3, They may be separated by up to 4, 5 or 10 weeks, up to 1, 2, 3, 4, 5 or 10 months or up to 1, 2, 3, 4, 5 or 10 years apart. In some embodiments, the kit of parts comprises a compound having a structure according to Formula (I), (II), (III) and/or (IV), or a compound of Table 1 above, or Formula (I), A pharmaceutical composition comprising a compound having a structure according to (II), (III) and/or (IV), or a compound of Table 1 above, and at least one pharmaceutically acceptable carrier include.

The composition may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, pH buffering agents and dispersing agents. Prevention of the presence of microorganisms can be ensured by sterilization procedures and/or by inclusion of various antibacterial and antifungal agents (eg, parabens, chlorobutanol, phenol, sorbic acid, etc.). It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical dosage form may be brought about by the inclusion of agents that slow absorption, such as aluminum monostearate and gelatin.

Regardless of the route of administration selected, the active compounds (which may be used in a suitable hydrated form) and/or pharmaceutical compositions used in accordance with the present invention are rendered pharmaceutically acceptable by conventional methods known to those of skill in the art. formulated into possible dosage forms (see, for example, Remington, "The Science and Practice of Pharmacy" edited by Allen, ^Loyd V., Jr., 22nd edition, Pharmaceutical Sciences, September 2012; Ansel et al., "Pharmaceutical Dosage Forms and Drug Delivery Systems", 7th edition, ^Lippincott Williams & Wilkins Publishers, 1999).

Pharmaceutical compositions can be administered by various methods known in the art. As will be appreciated by those skilled in the art, the route and/or mode of administration will vary depending on the desired result. Pharmaceutical compositions containing one or more active compounds employ carriers that will protect the one or more active compounds against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. can be prepared using Biodegradable, biocompatible polymers can be used, such as ethylene-vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparing such compositions are generally known to those skilled in the art. See, e.g., Sustained and Controlled Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

In order to administer a compound used in the invention by certain routes of administration, it may be necessary to coat the compound with, or co-administer the compound with, a material that will prevent its inactivation. For example, a compound can be administered to an individual in a suitable carrier such as a liposome or diluent. Pharmaceutically acceptable diluents include saline and aqueous buffer solutions. Liposomes include water-in-oil-in-water CGF emulsions, as well as conventional liposomes (Strejan et al., J. Neuroimmunol. 7: 27 (1984)).

Pharmaceutical compositions are generally sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyols (such as glycerol, propylene glycol and liquid polyethylene glycol), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by use of a coating such as lecithin, maintenance of the required particle size (in the case of dispersions) and use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of an injectable composition can be achieved by including in the composition a substance that delays absorption such as, for example, monostearate or gelatin.

An injectable composition should be sterile and fluid to the extent that the composition is deliverable by syringe. In addition to water, carriers can be isotonic buffered saline, ethanol, polyols such as glycerol, propylene glycol and liquid polyethylene glycol, and suitable mixtures thereof. Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterile microfiltration.

Dispersions generally are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying, which yield powders of the active ingredient plus, optionally, the desired ingredient from previously sterile-filtered solutions thereof. (lyophilized).

Dosage regimens are adjusted to provide the optimum desired response (eg, a therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally increased or decreased as indicated by the exigencies of the therapeutic condition. . It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the individual to be treated; each unit containing the desired pharmaceutical vehicle required. containing a predetermined amount of active compound calculated to produce a therapeutic effect of The specifications of the dosage unit forms used in accordance with the present invention include (a) the inherent characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the amount of such active compound for the treatment of sensitivities in individuals. dictated by, and directly dependent on, the limitations inherent in the art of compounding.

Examples of pharmaceutically acceptable antioxidants include (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium disulfite, sodium sulfite, etc.; (2) ascorbyl palmitate. , butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, Included are metal chelating agents such as tartaric acid, phosphoric acid, and the like.

For therapeutic/pharmaceutical formulations, the compositions used according to the invention are suitable for enteral administration (such as oral or rectal) or parenteral administration (such as nasal, topical (including vaginal, buccal and sublingual)). including The compositions may be conveniently presented in unit dosage form and prepared by any methods known in the art of pharmacy. The amount of active ingredient (particularly the amount of compound used according to the invention) that can be combined with a carrier material to produce a pharmaceutical composition (e.g., single dosage form) will depend on the individual to be treated and the particular administration. Varies depending on style. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will generally be that amount of the composition that produces a therapeutic effect.

Generally, out of 100% (for pharmaceutical formulations/compositions), the amount of active ingredient (especially the compounds used according to the invention (if present in pharmaceutical formulations/compositions, optionally combined with other therapeutically active agents) The amount of (a) ranges from about 0.01% to about 99%, preferably from about 0.1% to about 70%, most preferably from about 1% to about 30%, the remainder being Preferably, it consists of one or more pharmaceutically acceptable excipients.

The amount of an active ingredient, eg, a compound used in accordance with the invention, in unit dosage form and/or when administered to an individual or used in therapy is about 0.003 g per unit, dose or treatment. It may range from 1 mg to about 1000 mg (eg, from about 1 mg to about 500 mg, such as from about 10 mg to about 200 mg). In certain embodiments, the mass or body surface area of an individual (between about 1 mg/Kg and 10 mg/Kg (eg, between about 2 mg/Kg and 5 mg/Kg), or between about 1 mg/m ² and about 400 mg/m ² (e.g., between about 3 mg/m ² and about 350 mg/m ² , or between about 10 mg/m ² and about 200 mg/m ² ). A suitable amount may be calculated.

The actual dosage level of the active ingredient in the pharmaceutical compositions used in accordance with the present invention will be determined by providing the patient with an amount of the active ingredient effective to achieve the desired therapeutic response for a particular patient, composition and mode of administration. can be varied to obtain less toxic to The selected dosage level will be used in combination with the activity of the particular composition used, the route of administration, the time of administration, the excretion rate of the particular compound used, the duration of treatment, the particular composition used. Various pharmacokinetics, including other drugs, compounds and/or materials, the age, sex, weight, condition, general health and previous medical history of the patient being treated, and similar factors well known in the medical arts. Depends on strategic factors.

A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, a physician or veterinarian may begin with a dose of a compound used in accordance with the present invention at a level lower than that required to achieve the desired therapeutic effect, and increase the dose until the desired effect is achieved. It can be increased gradually. In general, a suitable daily dose of a composition for use in accordance with the invention will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such effective doses generally depend on the factors mentioned above. Administration is preferably oral, intravenous, intramuscular, intraperitoneal or subcutaneous, preferably administered proximal to the target site. If desired, the effective daily dose of the pharmaceutical composition as 2, 3, 4, 5, 6 or more sub-doses administered separately at appropriate intervals throughout the day, It may optionally be administered in unit dosage form. While it is possible for the compounds used according to the invention to be administered alone, it is preferable to administer the compounds as a pharmaceutical formulation/composition.

For oral administration, the pharmaceutical composition used according to the invention may contain, for example, binders (eg pregelatinized corn starch, polyvinylpyrrolidone, hydroxypropyl methylcellulose), fillers (eg lactose, microcrystalline cellulose, phosphorous). Calcium hydrogen phosphate), lubricants (e.g. magnesium stearate, talc, silica), disintegrants (e.g. potato starch, sodium starch glycolate) or wetting agents (e.g. sodium lauryl sulfate). It can take the form of tablets or capsules prepared by conventional means with the appropriate excipients. Liquid preparations for oral administration may be in the form of, for example, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations contain suspending agents (eg sorbitol, syrups, cellulose derivatives, hardened edible fats), emulsifiers (eg lecithin, gum arabic), non-aqueous vehicles (eg almond oil, oily esters, ethyl alcohol). , fractionated vegetable oils), preservatives (eg methyl or propyl-p-hydroxycarbonate, sorbic acid) and the like, with pharmaceutically acceptable additives, by conventional means. The preparations can also, if considered appropriate, contain buffer salts, flavoring, coloring and sweetening agents. Formulations for oral administration may be suitably formulated to give controlled release of the pharmaceutical composition of the invention.

In one embodiment, the compound is at most 100 mg/Kg body weight (e.g., at most 50 mg/Kg body weight, at most 40 mg/Kg body weight, at most 30 mg/Kg body weight, at most 20 mg/Kg body weight, at most 10 mg/Kg body weight). body weight, at most 5 mg/Kg body weight, at most 4 mg/Kg body weight, at most 3 mg/Kg body weight, at most 2 mg/Kg body weight, at most 1 mg/Kg body weight).

In one embodiment, the compound is at most 10 mg/Kg body weight (e.g., at most 5 mg/Kg body weight, at most 4 mg/Kg body weight, at most 3 mg/Kg body weight, at most 2 mg/Kg body weight, at most 1 mg/Kg body weight). body weight, at most 0.5 mg/Kg body weight, at most 0.4 mg/Kg body weight, at most 0.3 mg/Kg body weight, at most 0.2 mg/Kg body weight, at most 0.1 mg/Kg body weight) It is administered parenterally (eg intravenously, intramuscularly or subcutaneously).

The pharmaceutical composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate and the like.

A pharmaceutical composition for use according to the invention may be formulated for parenteral administration by injection, eg, by bolus injection or continuous infusion. In one embodiment, the compounds or compositions used according to the invention may be administered by slow continuous infusion over an extended period of time (eg, greater than 24 hours) to reduce toxic side effects. Administration may also be by continuous infusion over 2-24 hours, eg 2-12 hours. Such regimens may be repeated one or more times as necessary, eg, after 6 months or 12 months.

In yet another embodiment, the compounds or compositions used according to the invention are administered by maintenance therapy, eg once a week, eg for a period of 6 months or longer.

Formulations for injection can be presented in unit dosage form (eg, in vials, in multi-dose containers), with an added preservative. The pharmaceutical compositions for use according to the invention may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and contain formulatory agents such as suspending, stabilizing or dispersing agents. can be done. Alternatively, the drug may be in powder form for constitution with a suitable vehicle, eg, sterile pyrogen-free water, before use. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Optionally, the composition can also include a solubilizing agent and a local anesthetic (eg, lignocaine) to relieve pain at the injection site. Generally, the ingredients are supplied in unit dosage form either separately or mixed together, for example, as a lyophilized powder or anhydrous concentrate in a hermetically sealed container (such as an ampoule or sachette) indicating the quantity of active agent. be done. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients can be mixed prior to administration.

Compositions used according to the invention which are suitable for vaginal administration may also be pessaries, tampons, creams, gels, pastes, foams or pessaries containing such carriers as are known in the art to be suitable. Also includes spray formulations. Dosage forms for topical or transdermal administration of the compositions used according to the invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants that may be required.

In one embodiment, the compounds used according to the invention are formulated in liposomes. In a more preferred embodiment, the liposome contains a targeting moiety. In a most preferred embodiment, compounds in liposomes are delivered by bolus injection to a site proximal to the desired area. Such liposome-based compositions should be fluid to the extent that easy syringability exists, be stable under the conditions of manufacture and storage, and be susceptible to the contaminating action of microorganisms such as bacteria and fungi. should be preserved against

A "therapeutically effective dose" for therapy/treatment can be measured by objective responses, which can be either complete or partial. A complete response (CR) is defined as no clinical, radiological or other evidence of a condition, disorder or disease. A partial response (PR) is due to a disease reduction of greater than 50%. Median time to progression is a measure that characterizes the persistence of an objective tumor response.

A "therapeutically effective dose" for therapy/treatment can also be measured by its ability to stabilize the progression of a condition, disorder or disease. The ability of a compound to inhibit TRAF6 can be assessed using any of the in vitro or cell-based assays described herein that measure TRAF6 activity/inhibition. Alternatively, the properties of compounds described in this invention can be evaluated by testing the compound's potency in suitable animal model systems known to those of skill in the art. A therapeutically effective amount of a compound used in accordance with the present invention is intended to cure, cure, alleviate, alleviate, condition, disorder or disease, or a symptom of a condition, disorder or disease, or a predisposition to a condition, disorder or disease, in an individual. It can alleviate, alter, remedy, ameliorate, ameliorate or affect. One skilled in the art can determine such amounts based on factors such as the size of the individual, the severity of the individual's symptoms, and the particular composition or route of administration chosen.

The pharmaceutical compositions for use according to the invention may also be presented in a pack or dispenser device which may, if desired, contain one or more unit dosage forms containing the active compound. The pack can for example contain metal or plastic foil, such as a blister pack. The pack or dispenser device can be accompanied by instructions for administration.

The pharmaceutical compositions used according to the invention can be administered as the sole active agent or can be administered in combination with other therapeutic and/or cosmetic active agents. In one embodiment, the pharmaceutical composition used according to the invention is an antiviral agent, an antibody against an antigen of an animal pathogenic virus or another microorganism (e.g., pathogenic bacteria or fungi), or against a cancer antigen. agents that stimulate the subject's immune system (e.g., interferons such as interferon alpha or interferon beta, imiquimod and resiquimod), and one or more other therapeutically active agents selected from the group consisting of antimicrobial agents containing or administered with

Therapeutic and Other Uses In a further aspect, this application relates to compounds identified above under the heading "Compounds" or compounds identified above under the heading "Pharmaceutical Compositions" for use in therapy. A pharmaceutical composition is provided.

Generally, the present invention relates to compounds having structures according to formulas (I), (II), (III) and/or (IV), described herein under the heading "Compounds", or A compound or pharmaceutical composition identified above under the heading "Pharmaceutical Compositions" is capable of inhibiting or reducing the TRAF6-Ubc13 interaction, in particular inhibiting or reducing the enzymatic activity of TRAF6. Demonstrate what you can do.

Accordingly, in one aspect, the present invention provides compounds having structures according to formulas (I), (II), (III) and/or (IV), or compounds of Table 1, and hydrates, solvates thereof , salts, complexes, racemic mixtures, diastereomers, enantiomers and tautomers, and isotopically enriched forms of any of the foregoing for use in the treatment of TRAF6-related diseases, especially cancer, immune diseases , Parkinson's disease, cardiac hypertrophy or type 2 diabetes, preferably cancer, immune disorders and type 2 diabetes, more preferably cancer or immune disorders. Optionally, the method includes administering at least one additional active compound to the individual. At least one additional active compound is administered either before or after the compound having the structure of Formula (I), (II), (III) and/or (IV) or the compound of Table 1 can be done.

As used herein, throughout the description, the term "TRAF6-associated disease" refers to any pathological condition cumulatively caused by or correlated with TRAF6 overexpression or activity. or disease.

TRAF6 activity or overexpression has been shown to be associated with the following lesions:

Rheumatoid arthritis (RA): Disease initiation and progression of RA are significantly dependent on different signaling pathways of the innate and adaptive immune responses (McInnes, I.B., C.D. Buckley, and J.D. Isaacs, Cytokines in rheumatoid arthritis - shaping the immunological landscape. Nat Rev Rheumatol, 2016. 12(1): p. 63-8; McInnes, I.B. and G. Schett, Cytokines in the pathogenesis of rheumatoid arthritis. Nat Rev Immunol, 2007. 7(6): p. 429-42; Schett, G., J.M. Dayer, and B. Manger, Interleukin-1 function and role in rheumatic disease. Nat Rev Rheumatol, 2016. 12(1): p. 14-24; Choy, E., McInnes, I.B. and G. Schett, The pathogenesis of rheumatoid arthritis. N Engl J Med, 2011 365(23): p. 2205-19; Brzustewicz, E. and E. Bryl, The role of cytokines in the pathogenesis of rheumatoid arthritis--Practical and potential application of cytokines as biomarkers and targets of personalized therapy. Cytokine, 2015 76(2): p. 527-36 and Joosten, L.A., S. Abdollah. i-Roodsaz, C.A. Dinarello, et al., Toll-like receptors and chronic inflammation in rheumatic diseases: new developments. Nat Rev Rheumatol, 2016. 12(6): p. 344-57). Interestingly, all these pathways (IL-1β, TLRs, Th1, Th17 and others) are responsible for proper pathway activation, signal progression and cytokine (IL-1β, TNFα, IL-6 and others) secretion. use the TRAF6 E3 ligase activity. Furthermore, TRAF6 has been implicated in playing a role in osteoclast differentiation (Lamothe, B., W.K. Webster, A. Gopinathan, et al., TRAF6 ubiquitin ligase is essential for RANKL signaling and osteoclast differentiation). Biochem Biophys Res Commun, 2007. 359(4): p. 1044-9). Finally, studies on single nucleotide polymorphisms have linked TRAF6 to RA (Namjou, B., C.B. Choi, I.T. Harley, et al., Evaluation of TRAF6 in a large multiancestral lupus cohort. Arthritis Rheum, 2012. 64( 6): p. 1960-9). Inhibition of the TRAF6-Ubc13 interaction is therefore a novel strategy for reversing inflammation and bone resorption in RA. Our proof-of-concept studies in a CIA-induced preclinical RA mouse model substantiate this hypothesis that inhibition of the TRAF6-Ubc13 interaction by C25-140 (compound 3) indeed improves RA disease outcome. (See Figure 11). Moreover, TRAF6 expression was markedly elevated in joints of CIA-induced RA mice, indicating that siTRAF6 treatment of CIA-induced RA mice ameliorates arthritis index as well as inflammation and bone destruction (Wang, H., W. Chen Mol Med Rep, 2015. 11(4): p. 2761-6). Interestingly, the results of genetic loss of TRAF6 correlate perfectly with our TRAF6 activity inhibitor studies. Finally, a study observing 44 RA and 9 osteoarthritis patients revealed that TRAF6 expression was increased only in the RA patient pool, again suggesting that TRAF6 It may be predominantly involved in synovial inflammation and bone resorption through cell differentiation (Zhu, L.J., L. Dai, D.H. Zheng, et al., Upregulation of tumor necrosis factor receptor-associated factor 6 correlated with synovitis severity in rheumatoid arthritis. Arthritis Res Ther, 2012. 14(3): p. R133), thus underlining the argument that this may be an interesting target for the treatment of RA.

Psoriasis: The pathogenesis of psoriasis is highly dependent on IL-17 signaling, which includes TRAF6 as a key component in signal progression to cytokine expression (Baliwag, J., D.H. Barnes, and A. Johnston, Cytokines in psoriasis Cytokine, 2015. 73(2): p. 342-50; Grine, L., L. Dejager, C. Libert, et al., An inflammatory triangle in psoriasis: TNF, type I IFNs and IL-17. Growth Factor Rev, 2015. 26(1): p. 25-33 and Gu, C., L. Wu, and X. Li, IL-17 family: cytokines, receptors and signaling. Cytokine, 2013. 64(2) : p. 477-85). Furthermore, studies on TLR7 activation by imiquimod (IMQ) show that it drives psoriasis, thereby providing evidence that TLRs and the innate immune response are also involved in disease formation. (Gilliet, M., C. Conrad, M. Geiges, et al., Psoriasis triggered by toll-like receptor 7 agonist imiquimod in the presence of dermal plasmacytoid dendritic cell precursors. Arch Dermatol, 2004. 140(12): 1490-5). All TLRs, except TLR3, use TRAF6 as a central regulator of signaling. In line with this, the present application provides evidence from IMQ-induced psoriasis mouse studies that inhibition of the TRAF6-Ubc13 interaction by C25-140 improves disease outcome (see Figure 10).

Inflammatory Bowel Disease (IBD): Inflammation in IBD (Crohn's disease and ulcerative colitis) depends on K63 ubiquitination events promoted by TRAF6. It has been shown that the E3 ligase activity of TRAF6 is important for disease development (Watanabe, T., N. Asano, G. Meng, et al., NOD2 downregulates colonic inflammation by IRF4-mediated inhibition of K63-linked polyubiquitination). of RICK and TRAF6. Mucosal Immunol, 2014. 7(6): p. 1312-25 and Wei, J., C. Wei, M. Wang, et al., The GTPase-activating protein GIT2 protects against colitis by negatively regulating Toll-like receptor signaling. Proc Natl Acad Sci USA, 2014. 111(24): p. 8883-8). Moreover, recent studies have demonstrated that TLR2 signaling is involved in the development of colitis, as peptide inhibitors of the TLR2 receptor ameliorated DSS-induced colitis (Shmuel-Galia, L. EMBO J, 2016. 35(6): p. 685-98). In TLR2 signaling, TRAF6 is a key regulator of the progression of signals towards NF-κB and cytokine expression. Therefore, inhibitors of TRAF6 activity should be very suitable for the treatment of IBD. Moreover, clinical studies examining IBD patient samples found elevated levels of TRAF6 in both Crohn's disease and ulcerative colitis (Shen, J., Y. Qiao, Z. Ran, et al. , Different activation of TRAF4 and TRAF6 in inflammatory bowel disease. Mediators Inflamm, 2013. 2013: p. 647936), which again highlights the potential of targeting TRAF6 for IBD treatment.

Type 1 diabetes (T1D): IL-1-mediated NF-κB activation, which synergizes with TNFα and IFNγ, is a major driver of inflammation and apoptosis in β-cells (Bending, D., P. Zaccone , and A. Cooke, Inflammation and type one diabetes. Int Immunol, 2012. 24(6): p. 339-46; Csorba, T.R., A.W. Lyon, and M.D. Hollenberg, Autoimmunity and the pathogenesis of type 1 diabetes. Clin Lab Sci, 2010. 47(2): p. 51-71 and Mandrup-Poulsen, T., L. Pickersgill, and M.Y. Donath, Blockade of interleukin 1 in type 1 diabetes mellitus. Nat Rev Endocrinol, 2010. 6( 3): p. 158-66). Thus, inhibition of IL-1 signaling in animal models reduced T1D outcome and improved glycemia as well as β-cell function (Mandrup-Poulsen, T., L. Pickersgill, and M.Y. Donath, Blockade of interleukin 1 in type 1 diabetes mellitus. Nat Rev Endocrinol, 2010. 6(3): p. 158-66). To date, antibody-based IL-1 antagonists have been used in clinical trials and have shown efficacy against disease development (Baumann, B., H.H. Salem, and B.O. Boehm, Anti-inflammatory therapy in type 1 diabetes. Curr Diab Rep, 2012. 12(5): p.499-509). TLRs have also been shown to be involved in inflammation in insulitis (Eizirik, D.L., M.L. Colli, and F. Ortis, The role of inflammation in insulitis and beta-cell loss in type 1 diabetes. Nat Rev Endocrinol 2009. 5(4): 219-26). TRAF6 is a central regulator of IL-1 and TLR signaling, which may represent a novel strategy to reduce inflammation and prevent β-cell death in the pre-diabetic stage of insulitis .

Multiple sclerosis (MS): Previous studies have shown that experimental autoimmune encephalomyelitis (EAE), a mouse model for MS, is augmented by Th1 and Th17 immune responses (Jager, A. , V. Dardalhon, R.A. Sobel, et al., Th1, Th17, and Th9 effector cells induce experimental autoimmune encephalomyelitis with different pathological phenotypes. J Immunol, 2009. 183(11): p. 7169-77 and Zepp, J., L. Wu, and X. Li, IL-17 receptor signaling and T helper 17-mediated autoimmune demyelinating disease. Trends Immunol, 2011. 32(5): p. 232-9). Here IL-17 has been described as a major driver for the progression of MS (Zepp, J., L. Wu, and X. Li, IL-17 receptor signaling and T helper 17- mediated autoimmune demyelinating disease. Trends Immunol, 2011. 32(5): p. 232-9 and Petermann, F. and T. Korn, Cytokines and effector T cell subsets causing autoimmune CNS disease. FEBS Lett, 2011. 585(23) : p. 3747-57). TRAF6 activity also plays a central role in Th1 and Th17 signaling and may be an excellent target for MS treatment.

Celiac disease: Ingestion of wheat, barley or rye mediates inflammation of the small intestine in patients with celiac disease. Here, gluten induces an adaptive Th1 immune response in individuals with certain genetic predispositions. In addition, α-amylase/trypsin inhibitors (ATIs), pest resistance molecules in wheat, have been demonstrated to act as activators of the innate immune response. ATIs bind to the TLR4-MD2-CD14 complex and promote the activation and secretion of inflammatory cytokines (Junker, Y., S. Zeissig, S.J. Kim, et al., Wheat amylase trypsin inhibitors drive intestinal inflammation via activation of toll-like receptor 4. J Exp Med, 2012. 209(13): p. 2395-408). TRAF6 plays an important role in Th1 immune responses and TLR4 signaling and may serve as a novel target for the treatment of celiac disease.

Autoimmune diseases in general: Autoimmune diseases are caused by a combined elevation of immune and inflammatory signaling processes such as Th1, Th17, TLRs, IL-1 and TNF. Since TRAF6 is a key regulator in almost all of these signaling pathways that lead to NF-κB activation, inhibition of TRAF6 activity may be an effective strategy to target autoimmunity in general. very high. Cell-based experiments in the present application demonstrate that TRAF6-Ubc13 inhibitors interfere with these signaling processes.

Diffuse large B-cell lymphoma (DLBCL): DLBCL is subdivided into two major groups, the activated B-cell (ABC) type and the embryonic B-cell (GCB) type. ABC DLBCL survival depends on a variety of different mutations that lead to constitutively active NF-κB signaling. The major mutations are in B-cell receptor (BCR) and MYD88 signaling (Nagel, D., M. Vincendeau, A.C. Eitelhuber, et al., Mechanisms and consequences of constitutive NF-kappaB activation in B-cell lymphoid Oncogene, 2014. 33(50): p. 5655-65 and Ngo, V.N., R.M. Young, R. Schmitz, et al., Oncogenically active MYD88 mutations in human lymphoma. Nature, 2011. 470(7332): p 115-9). Current therapies mainly focus on BCR signaling, for example by compounds targeting BTK, MALT1, cIAP (Krappmann, D. and M. Vincendeau, Mechanisms of NF-kappaB deregulation in lymphoid malignancies. Semin Cancer Biol, 2016 and Wilson, W.H., Treatment strategies for aggressive lymphomas: what works? Hematology Am Soc Hematol Educ Program, 2013. 2013: p. 584-90). However, to date, IRAK4 inhibitors are the only available tools to reduce MYD88 signaling (Kelly, P.N., D.L. Romero, Y. Yang, et al., Selective interleukin-1 receptor-associated kinase 4 inhibitors for the treatment of autoimmune disorders and lymphoid malignancy. J Exp Med, 2015. 212(13): p. 2189-201). Our data in DLBCL with C25-140 (compound 3) demonstrate that this compound selectively kills ABC-DLBCL with chronic MYD88 signaling (see Figure 12). In line with this, Ubc13 inhibitors also successfully reduced NF-B signaling and survival of ABC-DLBCL cells (Pulvino, M., Y. Liang, D. Oleksyn, et al., Inhibition of proliferation and survival of diffuse large B-cell lymphoma cells by a small-molecule inhibitor of the ubiquitin-conjugating enzyme Ubc13-Uev1A. Blood, 2012. 120(8): p. 1668-77). However, inhibition of Ubc13 as an E2 enzyme has too broad an effect on cell signaling.

MALT Lymphoma: MALT lymphoma contains an API2-MALT1 fusion protein resulting from a chromosomal translocation. Among other things, this fusion protein cooperates with TRAF6 to initiate NF-κB activation leading to cancer development (Krappmann, D. and M. Vincendeau, Mechanisms of NF-kappaB deregulation in lymphoid malignancies. Semin Cancer Biol, 2016 and Staudt, L.M., Oncogenic activation of NF-kappaB. Cold Spring Harb Perspect Biol, 2010. 2(6): p.a000109). These pathways confer proliferation, survival and resistance to therapy. Therefore, inhibition of TRAF6 activity may be beneficial in MALT lymphoma.

Multiple myeloma (MM): A major driver of malignant growth of MM is intratumoral genomic instability and interactions between myeloma cells and the bone marrow microenvironment. Here, various signaling pathways are activated, including MAPK, JAK-STAT3, (PI3K)-AKT, NF-κB, Wnt, Notch, IGF and pleiotrophin signaling pathways, leading to secretion of cytokines and growth factors, Resulting in increased cell proliferation and osteoclast activity. Clinically, MM is characterized by osteolytic lesions, anemia, renal failure, hypercalcemia, peripheral neuropathy and others. However, bone disease is the most clinically important factor for morbidity. TRAF6 is involved in osteoclastogenesis in MM through its critical role in NF-κB activation by RANK signaling. Here, TRAF6 ligase activity in combination with Ubc13 is essential for signal progression. Therefore, inhibitors of TRAF6 activity have great potential to inhibit RANK signaling and thereby reduce bone metastasis and osteoclastogenesis in MM. Importantly, decoy peptides that cleave TRAF6 from the RANK receptor or siRNA against TRAF6 have already shown promising results in osteoclastogenesis and proliferation inhibition, respectively (Liu, H., S. Tamashiro, S. Baritaki, et al., TRAF6 activation in multiple myeloma: a potential therapeutic target. Clin Lymphoma Myeloma Leuk, 2012. 12(3): p. 155-63). Therefore, it is very likely that MM can be treated by inhibiting TRAF6.

Lung cancer: The 11p13 locus has been shown to be amplified in various lung cancers (NSCLC and SCLC lines). This amplification is associated with TRAF6 upregulation, thereby defining TRAF6 as an amplified oncogene in lung carcinoma. Amplified TRAF6 promotes Ras-driven activation of the proinflammatory transcription factor NF-κB and subsequent tumor-promoting responses (Starczynowski, D.T., W.W. Lockwood, S. Delehouzee, et al., TRAF6 is an amplified oncogene bridging). the RAS and NF-kappaB pathways in human lung cancer. J Clin Invest, 2011. 121(10): p. 4095-105). It has also been shown that the E3 ligase activity of TRAF6 is essential for EGFR-driven NF-κB activation and lung cancer progression (Pan, D., C. Jiang, Z. Ma, et al., MALT1 is required for EGFR-induced NF-kappaB activation and contributes to EGFR-driven lung cancer progression. Oncogene, 2016. 35(7): p. 919-28). Therefore, TRAF6 inhibition may be a good strategy for combination therapy in Ras-driven lung cancers, especially those that are resistant to current first-line therapies.

Lung Adenocarcinoma: The expression level of TRAF6 is significantly higher in lung adenocarcinoma cells, and TRAF6 is involved in the proliferation and invasion of latent lung adenocarcinoma cells (Zhong, L., F. Cao, and Q. You , Effect of TRAF6 on the biological behavior of human lung adenocarcinoma cell. Tumor Biol, 2013. 34(1): p. 231-9), which makes its inhibition relevant as a therapeutic approach for this type of cancer. suggesting that it is possible.

Colon cancer: TRAF6 has increased expression in colon cancer tissue when compared to normal lateral cancerous tissue. Expression levels of TRAF6 are negatively correlated with 5-year survival, which leads to decreased survival in patients with high levels of TRAF6 expression (Zhang, T., H. Wang, and L. Han, Expression and Clinical Significance of Tumor Necrosis Factor Receptor-Associated Factor 6 in Patients With Colon Cancer. Iran Red Crescent Med J, 2016. 18(1): p. e23931). This indicates that TRAF6 plays an important role in the development of colon cancer and that drugs targeting TRAF6 may have potential to treat this disease. Another study also demonstrated that TRAF6 expression is upregulated in colon cancer, which correlates with tumor grade/stage. RNAi experiments with siTRAF6 reduced colon cancer cell proliferation in combination with conventional anticancer agents 5-fluorouracil and etoposide treatment (Sun, H., X. Li, L. Fan, et al., TRAF6 is upregulated in colon cancer and promotes proliferation of colon cancer cells. Int J Biochem Cell Biol, 2014. 53: p. 195-201). Therefore, inhibition of TRAF6 may advance the effects of these drugs in combination treatments.

Prostate cancer: TRAF6 has been shown to ubiquitinate TGFβ type I receptor (TβRI) and presenilin 1 (PS1), promoting cleavage of TβRI's N-terminal intracellular domain (ICD) by TACE and PS1. The ICD then translocates to the nucleus where it acts as a transcriptional regulator of genes involved in tumor cell invasiveness. Importantly, the E3 ligase activity of TRAF6 is essential for this process (Gudey, S.K., R. Sundar, Y. Mu, et al., TRAF6 stimulates the tumor-promoting effects of TGFbeta type I receptor through polyubiquitination and activation). of presenilin 1. Sci Signal, 2014. 7(307): p. ra2 and Mu, Y., R. Sundar, N. Thakur, et al., TRAF6 ubiquitinates TGFbeta type I receptor to promote its cleavage and nuclear translocation in cancer Nat Commun, 2011. 2: p. 330). Furthermore, TGFβ induces MAPK signaling in a TRAF6-dependent manner to produce Snail1. This process induces invasiveness in aggressive prostate cancer (Thakur, N., S.K. Gudey, A. Marcusson, et al., TGFbeta-induced invasion of prostate cancer cells is promoted by c-Jun-dependent transcriptional activation of Snail1. Cell Cycle, 2014. 13(15): p.2400-14). Therefore, TRAF6 appears to be a good target in human prostate cancer.

Breast cancer: Recent studies have shown that TLR2 signaling promotes the development of breast cancer. Thus, TLR2 depletion or knockdown of its downstream regulators (MYD88 and IRAK1) reduced breast cancer growth (Scheeren, F.A., A.H. Kuo, L.J. van Weele, et al., A cell-intrinsic role for TLR2- MYD88 in intestinal and breast epithelia and oncogenesis. Nat Cell Biol, 2014. 16(12): p. 1238-48). Another study revealed that the TLR9-TRAF6 axis is significantly involved in breast cancer cell invasion. They also showed that samples from breast cancer patients contained elevated levels of TLR9 (Ilvesaro, J.M., M.A. Merrell, L. Li, et al., Toll-like receptor 9 mediates CpG oligonucleotide-induced cellular invasion Mol Cancer Res, 2008. 6(10): p. 1534-43). Both studies suggest that targeting TRAF6 as a key regulator of TLR2 and TLR9 signaling may be beneficial in breast cancer treatment.

Osteosarcoma: TRAF6 expression levels are significantly higher in osteosarcoma tissue. Moreover, expression levels were even higher when the osteosarcoma was based on lung metastases. This study could also demonstrate that TRAF6 positively regulates the proliferation and invasive potential of osteosarcoma cell lines (Meng, Q., M. Zheng, H. Liu, et al., TRAF6 regulates proliferation, apoptosis , and invasion of osteosarcoma cell. Mol Cell Biochem, 2012. 371(1-2): p. 177-86). Data in the present application show that C25-140 (Compound 3) reduces proliferation of U2OS cells to the same extent as 2 Gy irradiation and that C25-140 in combination with irradiation has a synergistic effect (see Figure 13). ). Therefore, TRAF6 inhibition represents a strategy to interfere with osteosarcoma in combination drug/radiotherapy.

Pancreatic cancer: In clinical trials, TRAF6 levels were shown to be upregulated in tissues of 53 pancreatic cancer patients. Moreover, overexpression of TRAF6 in pancreatic cancer cells induced cell proliferation and migration. In contrast, knockdown of TRAF6 using RNAi reduced the tumorigenicity of pancreatic cancer cells. More importantly, siRNA against TRAF6 reduced tumor growth in vivo (Rong, Y., D. Wang, W. Wu, et al., TRAF6 is over-expressed in pancreatic cancer and promotes the tumorigenicity of pancreatic cancer cells. Med Oncol, 2014. 31(11): p. 260.). In conclusion, TRAF6 presents a new target for developing pancreatic cancer therapeutics.

Esophageal squamous cell carcinoma (ESCC): TRAF6 expression is upregulated in ESCC based on a study analyzing 38 patient samples. Moreover, TRAF6 was increased in ESCC cell lines. In line with this, overexpression of TRAF6 promoted the proliferation of ESCC cells, whereas siRNA against TRAF6 showed adverse effects. Finally, siRNA against TRAF6 efficiently reduced tumor growth in vivo (Yao, F., Q. Han, C. Zhong, et al., TRAF6 promoted the tumorigenicity of esophageal squamous cell carcinoma. Tumor Biol, 2013. 34(5): p.3201-7). As a result, TRAF6 may present an attractive target for the development of ESCC therapeutics.

Parkinson's disease (PD): Chronic inflammation is thought to affect the development of PD (Block, M.L. and J.S. Hong, Microglia and inflammation-mediated neurodegeneration: multiple triggers with a common mechanism. Prog Neurobiol, 2005. 76(2) : pp. 77-98). Recent studies have shown that Parkin promotes proteasomal degradation of TRAF2 and TRAF6, preventing inflammatory responses and cytokine-induced cell death via the TNF and IL-1 pathways. In the case of Parkin loss or Parkin mutation, this protection is lost. Therefore, many human PD tissues show elevated TRAF6 levels (Chung, J.Y., H.R. Park, S.J. Lee, et al., Elevated TRAF2/6 expression in Parkinson's disease is caused by the loss of Parkin E3 ligase activity. Lab Invest , 2013. 93(6): p.663-76). Therefore, inhibition of TRAF6 may be a possible therapeutic strategy in Parkin-dependent PD patients.

Cardiac hypertrophy: Recent studies have demonstrated that TRAF6 levels are increased in pathological cardiac hypertrophy as a result of ROS production. More importantly, the authors demonstrate that the E3 ligase activity of TRAF6, which overactivates TAK1, is important for the cardiac hypertrophic response (Ji, Y.X., P. Zhang, X.J. Zhang, et al.). al., The ubiquitin E3 ligase TRAF6 exacerbates pathological cardiac hypertrophy via TAK1-dependent signaling. Nat Commun, 2016. 7: p. 11267). These data suggest that inhibition of TRAF6 activity may be a good opportunity to treat pathological cardiac hypertrophy.

Type 2 diabetes (T2D): Diet-induced obesity can promote inflammation and insulin resistance through IL-1β (Tack, C.J., R. Stienstra, L.A. Joosten, et al., Inflammation links excess fat to insulin). resistance: the role of the interleukin-1 family. Immunol Rev, 2012. 249(1): p. 239-52). Furthermore, it has been shown that monocytes from obese patients contain higher levels of TRAF6, thereby linking TRAF6 to metabolic disorders (Hulsmans, M., E. Van Dooren, C. Mathieu, et al., Decrease of miR-146b-5p in monocytes during obesity is associated with loss of the anti-inflammatory but not insulin signaling action of adiponectin. PLoS One, 2012. 7(2): p. e32794). Several other studies have further demonstrated the importance of TLR4 signaling for the development of insulin resistance (Pal, D., S. Dasgupta, R. Kundu, et al., Fetuin-A acts as an Nat Med, 2012. 18(8): p. 1279-85 and Uchimura, K., M. Hayata, T. Mizumoto, et al., The serine protease prostasin regulates endogenous ligand of TLR4 to promote lipid-induced insulin resistance. hepatic insulin sensitivity by modulating TLR4 signaling. Nat Commun, 2014. 5: p. 3428). Finally, mice deficient in CD40-TRAF6 signaling showed decreased body weight gain and decreased adipose tissue in a high-fat diet model (Chatzigeorgiou, A., T. Seijkens, B. Zarzycka, et al., Blocking CD40-TRAF6 signaling is a therapeutic target in obesity-associated insulin resistance. Proc Natl Acad Sci USA, 2014. 111(7): p. 2686-91). These studies collectively suggest that TRAF6 is a therapeutic target for obesity-induced insulin resistance. The data of the present application clearly demonstrate that C25-140 (compound 3) reduces body weight gain and also reduces IL-1β expression in the T2D mouse model (see Figure 9).

The terms "disease", "disorder" and "condition" are used interchangeably herein when used in the context of treatment or therapy (including prophylactic therapy) and any pathological condition ( cancer), in particular the pathological conditions (including cancerous forms) described herein.

In a further aspect, the present invention provides a pharmaceutical composition for use in the treatment of diseases associated with TRAF6, in particular cancer, immune disorders, Parkinson's disease, cardiac hypertrophy or type 2 diabetes, wherein the composition comprises formula (I) , (II), (III) and/or (IV), or a compound of Table 1 and one or more excipients and optionally at least one additional active compound. I will provide a. At least one additional active compound is administered either before or after the compound having the structure of Formula (I), (II), (III) and/or (IV) or the compound of Table 1 can be done.

In a further aspect, the invention provides a method of treating and/or preventing a condition, disorder or disease associated with TRAF6 overexpression, comprising administering a therapeutically effective amount of a compound described herein (especially a therapeutically effective amount of a compound having a structure of Formula (I), (II), (III) and/or (IV), or a compound of Table 1, to an individual in need thereof. do. Optionally, the method includes administering at least one additional active compound to the individual. at least one additional active compound is administered either before or after the compound having the structure of Formula (I), (II), (III) and/or (IV) or the compound of Table 1 can be done.

As used herein, the terms "subject" or "individual" are used interchangeably throughout the description and refer to animals (warm-blooded mammals such as humans and primates; birds; cats, dogs, sheep, Domestic or farm domesticated animals such as goats, cows, horses and pigs; laboratory animals such as mice, rats and guinea pigs; fish; reptiles; zoo and wild animals, etc.). The subject is preferably a mammal, more preferably a human.

As used herein, throughout the description, the term "effective amount" in the context of a composition or dosage form for administration to a subject includes cancer, immune disease, Parkinson's disease, cardiac hypertrophy or type 2 diabetes. Provide a therapeutic benefit to delay or minimize symptoms associated with cancer, immune disorders, Parkinson's disease, cardiac hypertrophy or type 2 diabetes, or cure or ameliorate cancer, immune disorders, Parkinson's disease, cardiac hypertrophy or type 2 diabetes It refers to the amount of a composition or dosage form sufficient to do so. Specifically, a therapeutically effective amount means an amount sufficient to provide a therapeutic benefit in vivo. When used in reference to an amount of a compound of the invention, the term preferably improves overall treatment, reduces or avoids symptoms or causes of disease, or enhances therapeutic efficacy of another therapeutic agent. or a non-toxic amount that enhances synergy with another therapeutic agent.

The effective amount will, of course, vary depending on the particular subject being treated; the severity of the condition, disease or disorder; individual patient parameters, including age, physical condition, size and weight; duration of treatment; Nature (if any); dependent on the particular route of administration and similar factors within the knowledge and expertise of the medical practitioner. These factors are well known to those skilled in the art and can be addressed with no more than routine experimentation. Generally, it is preferred that the maximum dose be used, ie, the highest safe dose according to sound medical judgment. However, it is understood by those skilled in the art that a patient may insist on a lower dose or a tolerable dose for medical reasons, psychological reasons or virtually any other reason.

In any of the above therapeutic aspects, the at least one additional active compound may be selected from the additional active compounds described above.

A method comprising administering a compound having a structure according to Formulas (I), (II), (III) and/or (IV), or a compound of Table 1 identified above, and adding at least one additional active compound In any of the above therapeutic modes involving administering, the at least one additional active compound may be formulated in a single pharmaceutical composition, together with the compound of the invention described above. Alternatively, the pharmaceutical composition comprises a compound having a structure according to Formulas (I), (II), (III) and/or (IV), or a compound of Table 1 is provided in a first formulation and at least one The additional active compound may be structured as a kit of parts provided in a second formulation, ie, a second pharmaceutical composition. The first and second pharmaceutical compositions may be combined prior to use. In other words, prior to administration of the pharmaceutical composition, formulations containing additional active compounds are added to compounds having structures according to formulas (I), (II), (III) and/or (IV), or It can be added to a first pharmaceutical composition containing the compound. Alternatively, the present teachings administer a compound having a structure according to Formula (I), (II), (III) and/or (IV), or a compound of Table 1, formulated into a first pharmaceutical composition and administering at least one additional active compound formulated in a second pharmaceutical composition. The pharmaceutical compositions may be administered simultaneously or sequentially. For example, a first pharmaceutical composition may be administered at a first time point and a second pharmaceutical composition may be administered at a second time point, which time points are, for example, 0, or 1, 2 , up to 3, 4, 5 or 10 minutes, up to 1, 2, 3, 4, 5 or 10 hours, up to 1, 2, 3, 4, 5 or 10 days, 1, 2, 3, 4, 5 or 10 It may be up to weeks, up to 1, 2, 3, 4, 5 or 10 months, or up to 1, 2, 3, 4, 5 or 10 months apart.

In some embodiments, the disease associated with TRAF6 is cancer, immune disease, Parkinson's disease, cardiac hypertrophy or type 2 diabetes, preferably cancer, immune disease and type 2 diabetes, more preferably cancer or immune disease. disease.

The cancer is preferably lymphoma such as diffuse large B cell lymphoma (DLBCL) and MALT lymphoma, multiple myeloma (MM), lung cancer, lung adenocarcinoma, colon cancer, prostate cancer, breast cancer, osteosarcoma, pancreas cancer and esophageal squamous cell carcinoma (ESCC)), in other embodiments, diffuse large B-cell lymphoma (DLBCL) and osteosarcoma.

The immune disease is preferably an autoimmune disease. Autoimmune diseases are preferably psoriasis, rheumatoid arthritis, celiac disease, inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, multiple sclerosis and diabetes type 1, in other embodiments psoriasis, rheumatoid arthritis and multiple Sclerosis, in another embodiment selected from the group consisting of psoriasis and rheumatoid arthritis.

As used herein, "cancer disease" or "cancer" includes diseases characterized by abnormally regulated growth, proliferation, differentiation, adhesion and/or migration of cells. By "cancer cell" is meant an abnormal cell that proliferates by rapid, uncontrolled cell proliferation and continues to proliferate after the stimulus that initiated the new growth ceases.

In general, the amount of a compound according to the invention described herein (especially a compound having a structure according to Formula (I), (II), (III) and/or (IV), or Table 1) is administered daily to an individual. 1) may be up to 100 mg/Kg (e.g., up to 50 mg/Kg, up to 40 mg/Kg, up to 30 mg/Kg, up to 20 mg/Kg, depending on factors such as the condition of the subject being treated and the mode of administration). Kg, up to 10 mg/Kg, up to 9 mg/Kg, up to 8 mg/Kg, up to 7 mg/Kg, up to 6 mg/Kg, up to 5 mg/Kg, up to 4 mg/Kg, up to 3 mg/Kg, up to 2 mg/Kg, or up to 1 mg /Kg). For example, an amount of a compound according to the invention described herein (particularly a compound having a structure according to Formula (I), (II), (III) and/or (IV), or Table 1) administered daily to an individual. The amount of compound of 1) will range from about 0.01 mg/Kg to 100 mg/Kg (e.g., from about 0.05 mg/Kg to 50 mg/Kg), depending on factors such as the condition of the subject being treated and the mode of administration. , about 0.1 mg/Kg to 40 mg/Kg, about 0.2 mg/Kg to 30 mg/Kg, about 0.3 mg/Kg to 20 mg/Kg, about 0.4 mg/Kg to 10 mg/Kg, about 0.5 mg/Kg to 9 mg/Kg, about 0.6 mg/Kg to 8 mg/Kg, about 0.7 mg/Kg to 7 mg/Kg, about 0.8 mg/Kg to 6 mg/Kg, about 0.8 mg/Kg to 6 mg/Kg. 9 mg/Kg to 5 mg/Kg, about 1 mg/Kg to 4 mg/Kg, or about 2 mg/Kg to 3 mg/Kg).

In one embodiment, a compound according to the invention described herein (particularly a compound having a structure according to formulas (I), (II), (III) and/or (IV) or compounds of Table 1) is up to 100 mg/Kg body weight (e.g. up to 50 mg/Kg body weight, up to 40 mg/Kg body weight, up to 30 mg/Kg body weight, up to 20 mg/Kg body weight, up to 10 mg/Kg body weight, up to 5 mg/Kg body weight, up to 4 mg/Kg body weight, It is administered orally at a concentration of up to 3 mg/Kg body weight, up to 2 mg/Kg body weight, up to 1 mg/Kg body weight).

In one embodiment, the compounds of the invention described herein (particularly compounds having a structure according to formulas (I), (II), (III) and/or (IV), or compounds of Table 1) are not more than 10 mg/Kg body weight (for example, not more than 5 mg/Kg body weight, not more than 4 mg/Kg body weight, not more than 3 mg/Kg body weight, not more than 2 mg/Kg body weight, not more than 1 mg/Kg body weight, not more than 0.5 mg/kg body weight) Kg body weight, at most 0.4 mg/Kg body weight, at most 0.3 mg/Kg body weight, at most 0.2 mg/Kg body weight, at most 0.1 mg/Kg body weight) administered parenterally (e.g., intravenously, intramuscularly or subcutaneously).

The embodiments and definitions of terms described in the context of the means of the invention, such as compounds according to formula (I), formula (II), formula (III), formula (IV) and/or Table 1, are the methods described above. and uses mutatis mutandis.

The present invention also provides a method of treating a disease associated with TRAF6 overexpression or activity in a subject, comprising an effective amount of formula (I), formula (II), formula (III), formula (IV) and /or A method comprising administering to said subject a compound listed in Table 1, or a pharmaceutically acceptable salt, solvate or hydrate thereof, or a pharmaceutical composition comprising said compound. Said method preferably further comprises administering at least one additional pharmaceutically active compound. The aspects, embodiments, definitions, etc. described above are also applicable to said method of treatment mutatis mutandis.

Furthermore, the present invention is explained in more detail by the following examples.

1) Materials and Methods 1.1) HPLC
The HPLC system consisted of an Agilent 1100 (Agilent , USA). Sample preparation: 1.0 mg of sample was dissolved in 0.8 mL of solvent A and 0.4 mL of solvent B and filtered through a 0.45 μm nylon membrane prior to analysis. Mobile phase: Solvent A was acetonitrile and Solvent B was 0.1% phosphoric acid in water.
Method 1: Gradient elution was programmed as follows: A/B = 5/95 (v/v, 0 min) → 95/5 (v/v, 4 min) → 95/5 (v/v, 5 .9 min) → 5/95 (v/v, 6.0 min). Column temperature: 25°C. Flow rate: 0.40 m/min. Detection wavelength: 254 nm. Injection volume: 2 μL.
Method 2: Gradient elution was programmed as follows: A/B = 20/80 (v/v, 0 min) → 95/5 (v/v, 3 min) → 95/5 (v/v, 5 .9 min) → 5/95 (v/v, 6.0 min). Column temperature: 35°C. Flow rate: 0.45 mL/min. Detection wavelength: 254 nm. Injection volume: 2 μL.

６－クロロ－３－メチル－［１，２，４］トリアゾロ［４，３－ｂ］ピリダジン（２．８０ｇ、１６．６mmol）のＥｔＯＨ（３０．０ｍＬ）溶液に、ヒドラジン水和物（８５％、２．９３mL、４９．８mmol）を加えた。反応混合物を８０℃で４時間加熱した。ＴＬＣ検査（ＥｔＯＡｃ、ＵＶ）は、出発物質（Ｒ_ｆ＝０．１０）が消費されたことを示した。反応物を２５℃に冷却した。沈殿物が生じ、２．５０ｇの所望の生成物が濾過後に得られた。
収率：２．５０ｇ（９１％）
¹H NMR: (400 MHz, DMSO-d₆): δ(ppm) = 7.84 (d, 1H), 6.75 (d, 1H), 4.28 (s, 2H), 2.52 (s, 3H)
参考文献：Polish Journal of Chemistry, 1996, 70, 10, 683-692 2) Preparation of Compound 2.1) 1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)hydrazine (P571-1)

To a solution of 6-chloro-3-methyl-[1,2,4]triazolo[4,3-b]pyridazine (2.80 g, 16.6 mmol) in EtOH (30.0 mL) was added hydrazine hydrate (85% , 2.93 mL, 49.8 mmol) was added. The reaction mixture was heated at 80° C. for 4 hours. A TLC check (EtOAc, UV) indicated that the starting material (R _f =0.10) was consumed. The reaction was cooled to 25°C. A precipitate formed and 2.50 g of the desired product was obtained after filtration.
Yield: 2.50 g (91%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 7.84 (d, 1H), 6.75 (d, 1H), 4.28 (s, 2H), 2.52 (s, 3H)
Reference: Polish Journal of Chemistry, 1996, 70, 10, 683-692

ＡｃＯＨ（１５．０mL）中のＰ５７１－１（１．５０ｇ、９．１４mmol）及びエチル ４－アセチル－５－オキソヘキサノアート（２．０１ｇ、１０．１mmol）の混合物を、１１０℃で４時間加熱した。ＴＬＣ検査（ＥｔＯＡｃ、ＵＶ）は、主スポットとして所望の生成物（Ｒ_ｆ＝０．２０）が観察できることを示した。反応物を濃縮してＡｃＯＨを除去した。残渣をＥｔＯＡｃで希釈し、５％ＮａＨＣＯ_３溶液、ブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させた。残渣をＥｔ_２Ｏから再結晶して、１．３０ｇの純粋な所望の生成物を得た。
収率： １．３０ｇ（４３％）
¹H NMR:(400 MHz, メタノール-d₄):δ(ppm) = 8.36 (d, 1H), 7.84 (d, 1H), 4.05 (q, 2H), 2.70 (t, 2H), 2.68 (s, 3H), 2.59 (s, 3H), 2.23 (s, 3H), 1.16 (t, 3H)
参考文献： European Journal of Medicinal Chemistry, 2013, 69, 701-710 2.2) 3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) Preparation of ethyl propanoate (P571-2)

A mixture of P571-1 (1.50 g, 9.14 mmol) and ethyl 4-acetyl-5-oxohexanoate (2.01 g, 10.1 mmol) in AcOH (15.0 mL) was heated at 110° C. for 4 hours. heated. TLC check (EtOAc, UV) showed desired product (R _f =0.20) observable as major spot. The reaction was concentrated to remove AcOH. The residue was diluted with EtOAc, washed with ₅ % _NaHCO3 solution, brine and dried over _Na2SO4 . The residue was recrystallized from Et ₂ O to give 1.30 g of pure desired product.
Yield: 1.30 g (43%)
¹ H NMR: (400 MHz, methanol-d ₄ ): δ(ppm) = 8.36 (d, 1H), 7.84 (d, 1H), 4.05 (q, 2H), 2.70 (t, 2H), 2.68 (s , 3H), 2.59 (s, 3H), 2.23 (s, 3H), 1.16 (t, 3H)
Reference: European Journal of Medicinal Chemistry, 2013, 69, 701-710

ＴＨＦ（１５．０mL）及び水（５．００mL）中のＰ５７１－２（１．３０ｇ、３．９６mmol）の溶液に、ＬｉＯＨ・Ｈ_２Ｏ（４９８mg、１１．９mmol）を加えた。反応混合物を２５℃で４時間撹拌した。ＴＬＣ検査（ＥｔＯＡｃ、ＵＶ）は、出発物質（Ｒ_ｆ＝０．２０）が消費され、所望の生成物が主スポット（Ｒ_ｆ＝０．０５）であることを示した。反応混合物を濃縮してＴＨＦを除去した。残渣を６ＮのＨＣｌでｐＨ＝３に調整し、沈殿物を得た。固体を濾過によって回収し、水で洗浄し、乾燥させ、９００mgの純粋な所望の生成物を得た。
収率：９００mg（７５％）
¹H NMR:(400 MHz, DMSO-d₆):δ(ppm) = 12.19 (s, 1H), 8.36 (d, 1H), 7.85 (d, 1H), 2.68 (s, 3H), 2.65 (d, 2H), 2.60 (s, 3H), 2.40 (t, 2H), 2.23 (s, 3H)
参考文献： European Journal of Medicinal Chemistry, 2013, 69, 701-710 2.3) 3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) Preparation of ethyl propanoate (P571-3)

To a solution of P571-2 (1.30 g, 3.96 mmol) in THF (15.0 mL) and water (5.00 mL) was added LiOH.H ₂ O (498 mg, 11.9 mmol). The reaction mixture was stirred at 25° C. for 4 hours. TLC check (EtOAc, UV) indicated starting material (R _f =0.20) was consumed and the desired product was the major spot (R _f =0.05). The reaction mixture was concentrated to remove THF. The residue was adjusted to pH=3 with 6N HCl to give a precipitate. The solid was collected by filtration, washed with water and dried to give 900 mg of pure desired product.
Yield: 900 mg (75%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 12.19 (s, 1H), 8.36 (d, 1H), 7.85 (d, 1H), 2.68 (s, 3H), 2.65 (d , 2H), 2.60 (s, 3H), 2.40 (t, 2H), 2.23 (s, 3H)
Reference: European Journal of Medicinal Chemistry, 2013, 69, 701-710

ＤＭＦ（３．００mL）中のＰ５７１－３（５０．０mg、０．１７mmol）、ＨＡＴＵ（６９．６mg、０．１８mmol）、１－ベンジルピペラジン（５８．７mg、０．３３mmol）及びＴＥＡ（３３．７mg、０．３３mmol）の混合物を、２５℃で１６時間撹拌した。ＴＬＣ検査（ＥｔＯＡｃ中１０％ＭｅＯＨ、ＵＶ）は、出発物質（Ｒ_ｆ＝０．１０）が消費され、所望の生成物が主スポット（Ｒ_ｆ＝０．３０）であることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（シリカゲル、ＥｔＯＡｃ中５０％ＭｅＯＨ）で精製して、５５．０mgの純粋な標的化合物を得た。
収率：５５．０mg（７２％）
¹H NMR:(400 MHz, DMSO-d₆):δ(ppm) = 8.38 (d, 1H), 7.86 (d, 1H), 7.30 - 7.25 (m, 1H), 7.23 - 7.20 (m, 2H), 3.44 (t, 2H), 3.39 (s, 2H), 2.66 (s, 3H), 2.63 (d, 2H), 2.58 (s, 3H), 2.46 (d, 2H), 2.26 (t, 2H), 2.22 (s, 3H), 2.17 (t, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４５９．２，Ｒｔ＝３．５７分（方法１） 2.4.1) 1-(4-benzylpiperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b] Preparation of pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one (HZM06-1)

P571-3 (50.0 mg, 0.17 mmol), HATU (69.6 mg, 0.18 mmol), 1-benzylpiperazine (58.7 mg, 0.33 mmol) and TEA (33.0 mg) in DMF (3.00 mL). 7 mg, 0.33 mmol) was stirred at 25° C. for 16 hours. A TLC check (10% MeOH in EtOAc, UV) indicated that the starting material (R _f =0.10) was consumed and the desired product was the major spot (R _f =0.30). The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phase was washed with water and brine, dried over Na ₂ SO ₄ and purified by chromatotron (silica gel, 50% MeOH in EtOAc) to give 55.0 mg of pure target compound.
Yield: 55.0 mg (72%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 8.38 (d, 1H), 7.86 (d, 1H), 7.30 - 7.25 (m, 1H), 7.23 - 7.20 (m, 2H) , 3.44 (t, 2H), 3.39 (s, 2H), 2.66 (s, 3H), 2.63 (d, 2H), 2.58 (s, 3H), 2.46 (d, 2H), 2.26 (t, 2H), 2.22 (s, 3H), 2.17 (t, 2H)
LC/MS: m/z = 459.2, Rt = 3.57 min (Method 1)

３－（３，５－ジメチル－１－（３－メチル－［１，２，４］－トリアゾロ－［４，３－ｂ］－ピリダジン－６－イル）－１Ｈ－ピラゾール－４－イル）－プロパン酸（５０mg、０．１７mmol）のジメチルホルムアミド（３mL）溶液を満たした丸底フラスコに、４－ベンジルピペリジン（０．０６mL、０．３３mmol）、ＨＡＴＵ（６８．５２mg、０．１８mmol）及びトリエチルアミン（０．０４mL、０．３０６mmol）を加えた。得られた反応混合物を、出発物質の完全な消費（１７時間）が観察されるまで、室温で撹拌した。次いで、水（１５mL）で希釈し、酢酸エチル（３×１０mL）で抽出した。合わせた有機層をＭｇＳＯ_４で乾燥させ、濾過し、真空中で濃縮した。生成物を、溶離剤として酢酸エチル／メタノール１００：００→５０：５０の直線勾配で溶離するフラッシュクロマトグラフィーによって精製した。適当な画分を合わせて、表記生成物５５mg（７０％）を得た。：¹H NMR (400 MHz, DMSO-d₆):δ8.4 (d, J = 10Hz, 1H), 7.88 (d, J = 10Hz, 1H), 7.33-7.09 (br m, 3H), 7.10-6.99 (br m, 2H), 4.39 (br d, 1H), 3.76 (br d, 1H), 2.86 (t, 1H), 2.71-2.61 (br m, 6H), 2.59 (s, 3H), 2.47-2.14 (br m, 4H), 2.24 (s, 3H), 1.77-1.63 (br m, 1H), 1.59-1.43 (br m, 2H), 0.99-0.73 (br m, 2H)；ＬＣ－ＭＳ（ＥＳＩ＋）ｍ／ｚ：［Ｍ＋Ｈ］^＋ Ｃ_２６Ｈ_３２Ｎ_７Ｏについての計算値：４５８．５８、実測値：４５８．６１、ｔ_Ｒ＝１．１６分。 2.4.2 1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]-triazolo-[4,3-b ]-pyridazin-6-yl)-1H-pyrazol-4-yl)-propan-1-one C25-140

3-(3,5-dimethyl-1-(3-methyl-[1,2,4]-triazolo-[4,3-b]-pyridazin-6-yl)-1H-pyrazol-4-yl)- 4-benzylpiperidine (0.06 mL, 0.33 mmol), HATU (68.52 mg, 0.18 mmol) and triethylamine were added to a round-bottomed flask filled with a solution of propanoic acid (50 mg, 0.17 mmol) in dimethylformamide (3 mL). (0.04 mL, 0.306 mmol) was added. The resulting reaction mixture was stirred at room temperature until complete consumption of starting material (17 hours) was observed. It was then diluted with water (15 mL) and extracted with ethyl acetate (3 x 10 mL). The combined organic layers _were dried over MgSO4, filtered and concentrated in vacuo. The product was purified by flash chromatography eluting with a linear gradient of ethyl acetate/methanol 100:00→50:50 as eluent. Appropriate fractions were combined to give 55 mg (70%) of the title product. : ¹ H NMR (400 MHz, DMSO-d ₆ ): 8.4 (d, J = 10Hz, 1H), 7.88 (d, J = 10Hz, 1H), 7.33-7.09 (br m, 3H), 7.10- 6.99 (br m, 2H), 4.39 (br d, 1H), 3.76 (br d, 1H), 2.86 (t, 1H), 2.71-2.61 (br m, 6H), 2.59 (s, 3H), 2.47- 2.14 (br m, 4H), 2.24 (s, 3H), 1.77-1.63 (br m, 1H), 1.59-1.43 (br m, 2H), 0.99-0.73 (br m, 2H); LC-MS (ESI+ ) m/z: [M ⁺ H] _calcd for _C26H32N7O : _458.58 , found: 458.61, _tR = 1.16 min.

ＤＭＦ（３．００mL）中のＰ５７１－３（５０．０mg、０．１７mmol）、ＨＡＴＵ（６９．６mg、０．１８mmol）、１－（３－メトキシベンジル）ピペラジン（６８．７mg、０．３３mmol）及びＴＥＡ（３３．７mg、０．３３mmol）の混合物を、２５℃で１６時間撹拌した。ＴＬＣ検査（ＥｔＯＡｃ中１０％ＭｅＯＨ、ＵＶ）は、出発物質（Ｒ_ｆ＝０．１０）が消費され、所望の生成物が主スポット（Ｒ_ｆ＝０．３０）であることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（シリカゲル、ＥｔＯＡｃ中５０％ＭｅＯＨ）で精製して、３０．０mgの純粋な標的化合物を得た。
収率：３０．０mg（３７％）
¹H NMR: (400 MHz, D₂O): δ(ppm) = 8.38 (d, 1H), 7.86 (d, 1H), 7.22 - 7.15 (m, 1H), 6.82 - 6.75 (m, 2H), 3.71 (s, 3H), 3.44 (d, 2H), 3.36 (s, 2H), 2.67 (s, 3H), 2.64 (d, 2H), 2.58 (s, 3H), 2.46 (d, 2H), 2.27 (s, 2H), 2.23 (s, 3H), 2.18 (d, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４８９．１，Ｒｔ＝３．６４分（方法１） 2.5) 1-(4-(3-methoxybenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3 Preparation of -b]pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one (HZM06-2)

P571-3 (50.0 mg, 0.17 mmol), HATU (69.6 mg, 0.18 mmol), 1-(3-methoxybenzyl)piperazine (68.7 mg, 0.33 mmol) in DMF (3.00 mL) and TEA (33.7 mg, 0.33 mmol) was stirred at 25° C. for 16 hours. A TLC check (10% MeOH in EtOAc, UV) indicated that the starting material (R _f =0.10) was consumed and the desired product was the major spot (R _f =0.30). The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phases were washed with water and brine, dried over Na ₂ SO ₄ and purified by chromatotron (silica gel, 50% MeOH in EtOAc) to give 30.0 mg of pure target compound.
Yield: 30.0 mg (37%)
¹ H NMR: (400 MHz, D ₂ O): δ(ppm) = 8.38 (d, 1H), 7.86 (d, 1H), 7.22 - 7.15 (m, 1H), 6.82 - 6.75 (m, 2H), 3.71 (s, 3H), 3.44 (d, 2H), 3.36 (s, 2H), 2.67 (s, 3H), 2.64 (d, 2H), 2.58 (s, 3H), 2.46 (d, 2H), 2.27 (s, 2H), 2.23 (s, 3H), 2.18 (d, 2H)
LC/MS: m/z = 489.1, Rt = 3.64 min (Method 1)

ＤＭＦ（３．００mL）中のＰ５７１－３（５０．０mg、０．１７mmol）、ＨＡＴＵ（６９．６mg、０．１８mmol）、１－（４－フルオロベンジル）ピペラジン（６４．７mg、０．３３mmol）及びＴＥＡ（３３．７mg、０．３３mmol）の混合物を、２５℃で１６時間撹拌した。ＴＬＣ検査（ＥｔＯＡｃ中１０％ＭｅＯＨ、ＵＶ）は、出発物質（Ｒ_ｆ＝０．１０）が消費され、所望の生成物が主スポット（Ｒ_ｆ＝０．３０）であることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（シリカゲル、ＥｔＯＡｃ中５０％ＭｅＯＨ）で精製して、５０．０mgの標的化合物を得た。
収率：５０．０mg（６３％）
¹H NMR: (400 MHz, DMSO-d₆): δ(ppm) = 8.38 (d, 1H), 7.85 (d, 1H), 7.28 - 7.23 (m, 1H), 7.13 - 7.08 (m, 2H), 3.45 (d, 2H), 3.38 (s, 2H), 2.67 (s, 3H), 2.63 (d, 2H), 2.58 (s, 3H), 2.46 (d, 2H), 2.26 (t, 2H), 2.22 (s, 3H), 2.18 (d, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４７７．１，Ｒｔ＝３．６４分（方法１） 2.6) 1-(4-(4-fluorobenzyl)piperazin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3 Preparation of -b]pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one (HZM06-3)

P571-3 (50.0 mg, 0.17 mmol), HATU (69.6 mg, 0.18 mmol), 1-(4-fluorobenzyl)piperazine (64.7 mg, 0.33 mmol) in DMF (3.00 mL) and TEA (33.7 mg, 0.33 mmol) was stirred at 25° C. for 16 hours. A TLC check (10% MeOH in EtOAc, UV) indicated that the starting material (R _f =0.10) was consumed and the desired product was the major spot (R _f =0.30). The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phase was washed with water and brine, dried over Na ₂ SO ₄ and purified by chromatotron (silica gel, 50% MeOH in EtOAc) to give 50.0 mg of target compound.
Yield: 50.0 mg (63%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 8.38 (d, 1H), 7.85 (d, 1H), 7.28 - 7.23 (m, 1H), 7.13 - 7.08 (m, 2H) , 3.45 (d, 2H), 3.38 (s, 2H), 2.67 (s, 3H), 2.63 (d, 2H), 2.58 (s, 3H), 2.46 (d, 2H), 2.26 (t, 2H), 2.22 (s, 3H), 2.18 (d, 2H)
LC/MS: m/z = 477.1, Rt = 3.64 min (Method 1)

ＤＭＡＣ（５．００mL）中のＰ５７１－３（２００mg、０．６７mmol）、ＨＡＴＵ（２５３mg、０．６７mmol）、３－ベンジルピロリジン（１０７mg、０．６７mmol）及びＴＥＡ（１３５mg、１．３３mmol）の混合物を、２５℃で１６時間撹拌した。ＨＰＬＣ－ＵＶ検査は、出発物質が消費され、所望の生成物が主要ピークであることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、塩基性Ａｌ_２Ｏ_３カラム（ＥｔＯＡｃ中１０％ＭｅＯＨ）で精製し、１００mgの純粋な標的化合物を得た。
収率：１００mg（３４％）
¹H NMR: (400 MHz, DMSO-d₆): δ(ppm) = 8.36 (dd, 1H), 7.84 (dd, 1H), 7.29 - 7.19 (m, 2H), 7.16 (t, 3H), 3.49 - 3.35 (m, 2H), 3.23-3.13 (m, 1H), 3.04 - 2.89 (m, 1H), 2.67 (d, 3H), 2.65-2.60 (m, 2H), 2.57 (d, 3H), 2.46-2.32 (m, 2H), 2.22 (d, 3H), 1.95-1.75 (m, 1H), 1.61 - 1.41 (m, 1H)
ＬＣ／ＭＳ：ｍ／ｚ＝４４４．３，Ｒｔ＝３．５５分（方法２） 2.7) 1-(3-benzylpyrrolidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- Preparation of 6-yl)-1H-pyrazol-4-yl)propan-1-one (HZM06-4)

A mixture of P571-3 (200 mg, 0.67 mmol), HATU (253 mg, 0.67 mmol), 3-benzylpyrrolidine (107 mg, 0.67 mmol) and TEA (135 mg, 1.33 mmol) in DMAC (5.00 mL) was stirred at 25° C. for 16 hours. HPLC-UV inspection indicated starting material was consumed and the desired product was the major peak. The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phase was washed with water and brine, dried over _Na2SO4 and purified _on a basic _Al2O3 column ( ₁₀ % MeOH in EtOAc) to give 100 mg of pure target compound.
Yield: 100 mg (34%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 8.36 (dd, 1H), 7.84 (dd, 1H), 7.29 - 7.19 (m, 2H), 7.16 (t, 3H), 3.49 - 3.35 (m, 2H), 3.23-3.13 (m, 1H), 3.04 - 2.89 (m, 1H), 2.67 (d, 3H), 2.65-2.60 (m, 2H), 2.57 (d, 3H), 2.46 -2.32 (m, 2H), 2.22 (d, 3H), 1.95-1.75 (m, 1H), 1.61 - 1.41 (m, 1H)
LC/MS: m/z = 444.3, Rt = 3.55 min (method 2).

ＤＭＦ（３．００mL）中のＰ５７１－３（５０．０mg、０．１７mmol）、ＨＡＴＵ（６９．６mg、０．１８mmol）、４－フェニルピペリジン（６４．７mg、０．３３mmol）及びＴＥＡ（３３．７mg、０．３３mmol）の混合物を、２５℃で１６時間撹拌した。ＴＬＣ検査（ＥｔＯＡｃ中１０％ＭｅＯＨ、ＵＶ）は、出発物質（Ｒ_ｆ＝０．１０）が消費され、所望の生成物が主スポット（Ｒ_ｆ＝０．３０）であることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（シリカゲル、ＥｔＯＡｃ中５０％ＭｅＯＨ）で精製し、４５．０mgの純粋な標的化合物を得た。
収率：４５．０mg（６１％）
¹H NMR:(400 MHz, DMSO-d₆): δ(ppm) = 8.36 (d, 1H), 7.84 (d, 1H), 7.15 - 7.02 (m, 5H), 4.57 (d, 1H), 3.90 (d, 1H), 3.03 (t, 1H), 2.69 - 2.64 (m, 2H), 2.62 (s, 3H), 2.60 (s, 3H), 2.47 - 2.37 (m, 1H), 2.26 (s, 3H), 1.71 (dd, 2H), 1.32-1.13 (m, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４４４．３，Ｒｔ＝４．５９分（方法１） 2.8) 3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) Preparation of -1-(4-phenylpiperidin-1-yl)propan-1-one (HZM06-6)

P571-3 (50.0 mg, 0.17 mmol), HATU (69.6 mg, 0.18 mmol), 4-phenylpiperidine (64.7 mg, 0.33 mmol) and TEA (33.0 mg, 0.18 mmol) in DMF (3.00 mL). 7 mg, 0.33 mmol) was stirred at 25° C. for 16 hours. A TLC check (10% MeOH in EtOAc, UV) indicated that the starting material (R _f =0.10) was consumed and the desired product was the major spot (R _f =0.30). The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phase was washed with water and brine, dried over Na ₂ SO ₄ and purified by chromatotron (silica gel, 50% MeOH in EtOAc) to give 45.0 mg of pure target compound.
Yield: 45.0 mg (61%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 8.36 (d, 1H), 7.84 (d, 1H), 7.15 - 7.02 (m, 5H), 4.57 (d, 1H), 3.90 (d, 1H), 3.03 (t, 1H), 2.69 - 2.64 (m, 2H), 2.62 (s, 3H), 2.60 (s, 3H), 2.47 - 2.37 (m, 1H), 2.26 (s, 3H) ), 1.71 (dd, 2H), 1.32-1.13 (m, 2H)
LC/MS: m/z = 444.3, Rt = 4.59 min (Method 1)

ＤＭＡＣ（３．００mL）中のＰ５７１－３（１２８mg、０．４３mmol）、ＨＡＴＵ（１７８mg、０．４７mmol）、２－（ピペリジン－４－イル）ピリジン二塩酸塩（１００mg、０．４３mmol）及びＴＥＡ（１７２mg、１．７０mmol）の混合物を、２５℃で１６時間撹拌した。ＨＰＬＣ検査は、出発物質が消費され、所望の生成物が主要ピークであることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（塩基性Ａｌ_２Ｏ_３、ＥｔＯＡｃ中５０％ＭｅＯＨ）によって精製して、３７．０mgの純粋な標的化合物を得た。
収率：３７．０mg（２０％）
¹H NMR: (400 MHz, DMSO-d₆): δ(ppm) = 8.34 (d, 1H), 8.32 (d, 1H), 7.82 (d, 1H), 7.56 (td, 1H), 7.14 (d, 1H), 7.09 (ddd, 1H), 4.55 (d, 1H), 3.90 (d, 1H), 3.06 (t, 1H), 2.73 - 2.65 (m, 2H), 2.63 (s, 3H), 2.59 (s, 3H), 2.25 (s, 3H), 1.80 (dd, 2H), 1.47 - 1.23 (m, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４４５．２，Ｒｔ＝３．４４分（方法１） 2.9) 3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) Preparation of -1-(4-(pyridin-2-yl)piperidin-1-yl)propan-1-one (HZM06-7)

P571-3 (128 mg, 0.43 mmol), HATU (178 mg, 0.47 mmol), 2-(piperidin-4-yl)pyridine dihydrochloride (100 mg, 0.43 mmol) and TEA in DMAC (3.00 mL) A mixture of (172 mg, 1.70 mmol) was stirred at 25° C. for 16 hours. HPLC check showed the starting material to be consumed and the desired product to be the major peak. The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phases were washed with water and brine, dried over Na ₂ SO ₄ and purified by chromatotron (basic Al ₂ O ₃ , 50% MeOH in EtOAc) to give 37.0 mg of pure target compound. Obtained.
Yield: 37.0 mg (20%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 8.34 (d, 1H), 8.32 (d, 1H), 7.82 (d, 1H), 7.56 (td, 1H), 7.14 (d , 1H), 7.09 (ddd, 1H), 4.55 (d, 1H), 3.90 (d, 1H), 3.06 (t, 1H), 2.73 - 2.65 (m, 2H), 2.63 (s, 3H), 2.59 ( s, 3H), 2.25 (s, 3H), 1.80 (dd, 2H), 1.47 - 1.23 (m, 2H)
LC/MS: m/z = 445.2, Rt = 3.44 min (Method 1)

ＤＭＦ（３．００mL）中のＰ５７１－３（５０．０mg、０．１７mmol）、ＨＡＴＵ（６９．６mg、０．１８mmol）、４－（３－メトキシベンジル）ピペリジン塩酸塩（８０．０mg、０．３３mmol）及びＴＥＡ（６７．４mg、０．６７mmol）の混合物を、２５℃で１６時間撹拌した。ＴＬＣ検査（ＥｔＯＡｃ中１０％ＭｅＯＨ、ＵＶ）は、出発物質（Ｒ_ｆ＝０．１０）が消費され、所望の生成物が主スポット（Ｒ_ｆ＝０．３０）であることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（シリカゲル、ＥｔＯＡｃ中５０％ＭｅＯＨ）で精製し、続いてＥｔ_２Ｏから再結晶し、３０．０mgの純粋な標的化合物を得た。
収率：３０．０mg（３７％）
¹H NMR:(400 MHz, D₂O): δ(ppm) = 8.38 (d, 1H), 7.87 (d, 1H), 7.11 (t, 1H), 6.74 - 6.69 (m, 1H), 6.65 - 6.57 (m, 2H), 4.38 (d, 1H), 3.76 (d, 1H), 3.69 (s, 3H), 2.85 (t, 1H), 2.67 (s, 3H), 2.59 (s, 3H), 2.47 - 2.38 (m, 2H), 2.23 (s, 3H), 1.69 (s, 1H), 1.51 (m, 2H), 0.99 - 0.70 (m, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４８８．２，Ｒｔ＝４．８７分（方法１） 2.10) 1-(4-(3-methoxybenzyl)piperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3 Preparation of -b]pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one (HZM06-8)

P571-3 (50.0 mg, 0.17 mmol), HATU (69.6 mg, 0.18 mmol), 4-(3-methoxybenzyl)piperidine hydrochloride (80.0 mg, 0.18 mmol) in DMF (3.00 mL). 33 mmol) and TEA (67.4 mg, 0.67 mmol) was stirred at 25° C. for 16 hours. A TLC check (10% MeOH in EtOAc, UV) indicated that the starting material (R _f =0.10) was consumed and the desired product was the major spot (R _f =0.30). The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phases were washed with water and brine, dried over Na ₂ SO ₄ and purified by chromatotron (silica gel, 50% MeOH in EtOAc) followed by recrystallization from Et ₂ O to give 30.0 mg of pure We obtained a suitable target compound.
Yield: 30.0 mg (37%)
¹ H NMR: (400 MHz, D ₂ O): δ(ppm) = 8.38 (d, 1H), 7.87 (d, 1H), 7.11 (t, 1H), 6.74 - 6.69 (m, 1H), 6.65 - 6.57 (m, 2H), 4.38 (d, 1H), 3.76 (d, 1H), 3.69 (s, 3H), 2.85 (t, 1H), 2.67 (s, 3H), 2.59 (s, 3H), 2.47 - 2.38 (m, 2H), 2.23 (s, 3H), 1.69 (s, 1H), 1.51 (m, 2H), 0.99 - 0.70 (m, 2H)
LC/MS: m/z = 488.2, Rt = 4.87 min (Method 1)

ＤＭＡＣ（３．００mL）中のＰ５７１－３（１００mg、０．３３mmol）、ＨＡＴＵ（１３３mg、０．３５mmol）、４－（４－フルオロベンジル）ピペリジン（６４．０mg、０．３３mmol）及びＴＥＡ（６７．４mg、０．６７mmol）の混合物を、２５℃で１６時間撹拌した。ＴＬＣ検査（ＥｔＯＡｃ中１０％ＭｅＯＨ、ＵＶ）は、出発物質（Ｒ_ｆ＝０．１０）が消費され、所望の生成物が主スポット（Ｒ_ｆ＝０．３０）であることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（シリカゲル、ＥｔＯＡｃ中５０％ＭｅＯＨ）で精製し、４５．０mgの純粋な標的化合物を得た。
収率：４５．０mg（２８％）
¹H NMR: (400 MHz, DMSO-d₆): δ(ppm) = 8.36 (d, 1H), 7.84 (d, 1H), 7.14 - 7.05 (m, 2H), 7.04-6.96 (m, 2H), 4.36 (d, 1H), 3.75 (d, 1H), 2.83 (t, 1H), 2.65 (s, 3H), 2.57 (s, 3H), 2.41 - 2.36 (m, 2H), 2.21 (s, 3H), 1.72 - 1.58 (m, 1H), 1.48 (dd, 2H), 0.98 - 0.72 (m, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４７６．１，Ｒｔ＝４．９７分（方法１） 2.11) 1-(4-(4-fluorobenzyl)piperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3 Preparation of b]pyridazin-6-yl)-1H-pyrazol-4-yl)propan-1-one (HZM06-9)

P571-3 (100 mg, 0.33 mmol), HATU (133 mg, 0.35 mmol), 4-(4-fluorobenzyl)piperidine (64.0 mg, 0.33 mmol) and TEA (67 mg) in DMAC (3.00 mL) .4 mg, 0.67 mmol) was stirred at 25° C. for 16 hours. A TLC check (10% MeOH in EtOAc, UV) indicated that the starting material (R _f =0.10) was consumed and the desired product was the major spot (R _f =0.30). The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phase was washed with water and brine, dried over Na ₂ SO ₄ and purified by chromatotron (silica gel, 50% MeOH in EtOAc) to give 45.0 mg of pure target compound.
Yield: 45.0 mg (28%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 8.36 (d, 1H), 7.84 (d, 1H), 7.14 - 7.05 (m, 2H), 7.04-6.96 (m, 2H) , 4.36 (d, 1H), 3.75 (d, 1H), 2.83 (t, 1H), 2.65 (s, 3H), 2.57 (s, 3H), 2.41 - 2.36 (m, 2H), 2.21 (s, 3H ), 1.72 - 1.58 (m, 1H), 1.48 (dd, 2H), 0.98 - 0.72 (m, 2H)
LC/MS: m/z = 476.1, Rt = 4.97 min (Method 1)

ＤＭＡＣ（３．００mL）中のＰ５７１－３（１２０mg、０．４０mmol）、ＨＡＴＵ（１６０mg、０．４２mmol）、２－（（ピペリジン－４－イル）メチル）ピリジン二塩酸塩（１００mg、０．４０mmol）及びＴＥＡ（１６２mg、１．６１mmol）の混合物を、２５℃で１６時間撹拌した。ＨＰＬＣ検査は、出発物質が消費され、所望の生成物が主要ピークであることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（塩基性Ａｌ_２Ｏ_３、ＥｔＯＡｃ中５０％ＭｅＯＨ）によって精製し、５０．０mgの純粋な標的化合物を得た。
収率：５０．０mg（２７％）
¹H NMR: (400 MHz, メタノール-d₄): δ(ppm) = 8.41 (dt, 1H), 8.37 (d, 1H), 7.86 (d, 1H), 7.62 (td, 1H), 7.16 (ddd, 1H), 7.09 (d, 1H), 4.37 (d, 1H), 3.76 (d, 1H), 2.86 (q, 1H), 2.67 (s, 3H), 2.65 - 2.61 (m, 2H), 2.58 (s, 3H), 2.54 (dd, 2H), 2.47 - 2.35 (m, 2H), 2.23 (s, 3H), 1.50 (dd, 2H), 1.10-0.80 (m, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４５９．２，Ｒｔ＝３．４９分（方法１） 2.12) 3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) Preparation of -1-((4-(pyridin-2-yl)methyl)piperidin-1-yl)propan-1-one (HZM06-10)

P571-3 (120 mg, 0.40 mmol), HATU (160 mg, 0.42 mmol), 2-((piperidin-4-yl)methyl)pyridine dihydrochloride (100 mg, 0.40 mmol) in DMAC (3.00 mL) ) and TEA (162 mg, 1.61 mmol) was stirred at 25° C. for 16 h. HPLC check showed the starting material to be consumed and the desired product to be the major peak. The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phases were washed with water and brine, dried over _Na2SO4 and purified by chromatotron ₍ basic _{Al2O3 ,} 50% MeOH in EtOAc) to give 50.0 mg of pure target compound. rice field.
Yield: 50.0 mg (27%)
¹ H NMR: (400 MHz, methanol-d ₄ ): δ(ppm) = 8.41 (dt, 1H), 8.37 (d, 1H), 7.86 (d, 1H), 7.62 (td, 1H), 7.16 (ddd , 1H), 7.09 (d, 1H), 4.37 (d, 1H), 3.76 (d, 1H), 2.86 (q, 1H), 2.67 (s, 3H), 2.65 - 2.61 (m, 2H), 2.58 ( s, 3H), 2.54 (dd, 2H), 2.47 - 2.35 (m, 2H), 2.23 (s, 3H), 1.50 (dd, 2H), 1.10-0.80 (m, 2H)
LC/MS: m/z = 459.2, Rt = 3.49 min (Method 1)

ＤＭＦ（３．００mL）中のＰ５７１－３（５０．０mg、０．１７mmol）、ＨＡＴＵ（６９．６mg、０．１８mmol）、１－（（ピリジン－２－イル）メチル）ピペラジン（５９．０mg、０．３３mmol）及びＴＥＡ（３３．７mg、０．３３mmol）の混合物を、２５℃で１６時間撹拌した。ＨＰＬＣ検査は、出発物質が消費され、所望の生成物が主要ピークであることを示した。反応混合物を水（１５．０mL）で希釈し、ＥｔＯＡｃ（１０．０mL×５）で抽出した。合わせた有機相を水及びブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、クロマトトロン（シリカゲル、ＭｅＯＨ）で精製し、２５．０mgの純粋な標的化合物を得た。
収率：２５．０mg（３３％）
¹H NMR: (400 MHz, DMSO-d₆): δ(ppm) = 8.43 (dt, 1H), 8.37 (d, 1H), 7.85 (d, 1H), 7.73 (td, 1H), 7.41 - 7.37 (m, 1H), 7.23 (ddd, 1H), 3.52 (s, 2H), 3.46 (t, 2H), 3.39 (d, 2H), 2.67 (s, 3H), 2.64 (d, 2H), 2.58 (s, 3H), 2.47 (d, 2H), 2.33 (t, 2H), 2.26 (t, 2H), 2.23 (s, 3H)
ＬＣ／ＭＳ：ｍ／ｚ＝４６０．２，Ｒｔ＝３．４８分（方法１） 2.13) 3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-6-yl)-1H-pyrazol-4-yl) Preparation of -1-((4-(pyridin-2-yl)methyl)piperazin-1-yl)propan-1-one (HZM06-11)

P571-3 (50.0 mg, 0.17 mmol), HATU (69.6 mg, 0.18 mmol), 1-((pyridin-2-yl)methyl)piperazine (59.0 mg, 0.33 mmol) and TEA (33.7 mg, 0.33 mmol) was stirred at 25° C. for 16 hours. HPLC check showed the starting material to be consumed and the desired product to be the major peak. The reaction mixture was diluted with water (15.0 mL) and extracted with EtOAc (10.0 mL x 5). The combined organic phase was washed with water and brine, dried over _Na2SO4 and purified by chromatotron ₍ silica gel, MeOH) to give 25.0 mg of pure target compound.
Yield: 25.0 mg (33%)
¹ H NMR: (400 MHz, DMSO-d ₆ ): δ(ppm) = 8.43 (dt, 1H), 8.37 (d, 1H), 7.85 (d, 1H), 7.73 (td, 1H), 7.41 - 7.37 (m, 1H), 7.23 (ddd, 1H), 3.52 (s, 2H), 3.46 (t, 2H), 3.39 (d, 2H), 2.67 (s, 3H), 2.64 (d, 2H), 2.58 ( s, 3H), 2.47 (d, 2H), 2.33 (t, 2H), 2.26 (t, 2H), 2.23 (s, 3H)
LC/MS: m/z = 460.2, Rt = 3.48 min (Method 1)

ＬＣ－ＭＳ方法３：Ａｑｕｉｔｙ ＵＰＬＣ ＢＥＨ Ｃ１８カラム（１．７μm、２．１×５０mm）を備えたＷａｔｅｒｓ Ａｃｑｕｉｔｙ Ｈ ＵＰＬＣ ＣＬＡＳＳシステム、流速０．８mL／分、水／アセトニトリル（それぞれに０．０５％ｖ／ｖＴＦＡを添加）からなる移動相を用いた３分間勾配法を使用：９５：５→５：９５（０～２．２５分）、９５：５（２．２７～３分）。
ＬＣ－ＭＳ方法４（Ａｇｉｌｅｎｔ １２００ Ｓｅｒｉｅｓ ＬＣシステム；Ａｇｉｌｅｎｔ ６１３０ Ｑｕａｄｒｕｐｏｌｅ検出器；カラム：ＬＵＮＡ Ｃ１８（２）カラム（３μm、１０×２mm））：
流量：１．１mL／分
０．０－０．２分 ７％ ＣＨ_３ＣＮ
０．２－１．８分 ７→９５％ ＣＨ_３ＣＮ
１．８－１．９分 ９５％ ＣＨ_３ＣＮ
１．９－２．０分 ９５→７％ ＣＨ_３ＣＮ The following compounds were obtained following synthetic procedures analogous to those described for the examples:

LC-MS Method 3: Waters Acquity H UPLC CLASS system equipped with an Aquity UPLC BEH C18 column (1.7 μm, 2.1×50 mm), flow rate 0.8 mL/min, water/acetonitrile (0.05% v /vTFA added) using a 3 min gradient method with a mobile phase consisting of: 95:5→5:95 (0-2.25 min), 95:5 (2.27-3 min).
LC-MS method 4 (Agilent 1200 Series LC system; Agilent 6130 Quadrupole detector; Column: LUNA C18(2) column (3 μm, 10×2 mm)):
Flow rate: 1.1 mL/min 0.0-0.2 min 7% CH ₃ CN
0.2-1.8 min 7→95% CH ₃ CN
1.8-1.9 min 95% CH ₃ CN
1.9-2.0 min 95→7% CH ₃ CN

実施例 Ｃ２５－１４０をトルエンに溶解し、ローソン試薬（０．６当量）を加えた。反応混合物を封管中、１１０℃で３時間撹拌した。粗混合物を真空中で濃縮し、シリカ上でのカラムクロマトグラフィー（ＣＨ_２Ｃｌ_２／ｉＰｒＯＨ）により精製して、所望の生成物（収率９６％）を得た。
¹H NMR: (400 MHz, CDCl₃): δ(ppm) = 8.08 (d, 1H, J = 9.9 Hz), 7.98 (d, 1H, J = 9.9 Hz), 7.21-7.25 (m, 2H), 7.16-7.21 (m, 1H), 7.03-7.07 (m, 2H), 5.67 (m, 1H), 4.14 (m, 1H), 3.05 (ddd, 1H, J = 12.5, 12.5, 2.0 Hz), 2.85-3.00 (m, 5H), 2.76 (s, 3H), 2.66 (s, 3H), 2.57 (dd, 1H, J = 13.5, 6.7 Hz), 2.44 (dd, 1H, J = 13.3, 7.5 Hz), 2.29 (s, 3H), 1.67-1.91 (m, 3H), 1.35 (dddd, 1H, J = 12.5, 12.5, 12.5, 3.3 Hz), 1.03 (dddd, 1H, J = 12.3, 12.3, 12.3, 4.0 Hz)
ＬＣ／ＭＳ： ｍ／ｚ＝４７３．９、Ｒ_ｔ＝１．５５分（方法４） 2.14) 1-(4-benzylpiperidin-1-yl)-3-(3,5-dimethyl-1-(3-methyl-[1,2,4]triazolo[4,3-b]pyridazine- Preparation of 6-yl]-1H-pyrazol-4-yl)propane-1-thione (45)

Example C25-140 was dissolved in toluene and Lawesson's reagent (0.6 eq) was added. The reaction mixture was stirred at 110° C. for 3 hours in a sealed tube. The crude mixture was concentrated in vacuo and purified by column chromatography on silica (CH ₂ Cl ₂ /iPrOH) to give the desired product (96% yield).
¹ H NMR: (400 MHz, CDCl ₃ ): δ(ppm) = 8.08 (d, 1H, J = 9.9 Hz), 7.98 (d, 1H, J = 9.9 Hz), 7.21-7.25 (m, 2H), 7.16-7.21 (m, 1H), 7.03-7.07 (m, 2H), 5.67 (m, 1H), 4.14 (m, 1H), 3.05 (ddd, 1H, J = 12.5, 12.5, 2.0 Hz), 2.85- 3.00 (m, 5H), 2.76 (s, 3H), 2.66 (s, 3H), 2.57 (dd, 1H, J = 13.5, 6.7Hz), 2.44 (dd, 1H, J = 13.3, 7.5Hz), 2.29 (s, 3H), 1.67-1.91 (m, 3H), 1.35 (dddd, 1H, J = 12.5, 12.5, 12.5, 3.3 Hz), 1.03 (dddd, 1H, J = 12.3, 12.3, 12.3, 4.0 Hz)
LC/MS: m/z = 473.9, _Rt = 1.55 min (Method 4).

チオアミド４５のＴＨＦ溶液に、ＭｅＩ（５０当量）を加えた。転化が完了するまで反応物を室温で撹拌し、溶媒を真空中で除去した。粗混合物を酢酸エチル及び水（１：１）に溶解し、ＮａＢＨ_３ＣＮ（１．１当量）を加えた。反応が完了するまで混合物を撹拌し、２０％水酸化ナトリウム水溶液を添加してクエンチし、ジエチルエーテルで数回抽出した。合わせた有機相をＮａ_２ＳＯ_４で乾燥し、溶媒を真空中で除去し、粗生成物をシリカでのカラムクロマトグラフィー（ＣＨ_２Ｃｌ_２／ＭｅＯＨ）により精製して、所望の生成物（４１％）を得た。
¹H NMR: (400 MHz, CDCl₃): δ(ppm) = 8.06 (d, 1H, 10.2 Hz), 7.99 (d, 1H, J = 9.9 Hz), 7.24-7.29 (m, 2H), 7.16-7.21 (m, 1H), 7.11-7.15 (m, 2H) 2.97 (d, 1H, J = 12.3 Hz), 2.77 (s, 3H), 2.63 (s, 3H), 2.54 (d, 1H, J = 6.8 Hz), 2.37-2.47 (m, 5H), 2.26 (s, 3H), 1.95 (t, 2H, J = 11.3 Hz), 1.62-1.78 (m, 4H), 1.48-1.61 (m, 1H), 1.34-1.44 (m, 2H)
ＬＣ／ＭＳ：ｍ／ｚ＝４４４．０、Ｒ_ｔ＝１．１６分（方法４） 2.15) 6-(4-(3-(4-benzylpiperidin-1-yl)-3,5-dimethyl-1H-pyrazol-1-yl)-3-methyl-[1,2,4]triazolo Preparation of [4,3-b]pyridazine (46)

To a THF solution of thioamide 45 was added MeI (50 eq). The reaction was stirred at room temperature until conversion was complete and the solvent was removed in vacuo. The crude mixture was dissolved in ethyl acetate and water (1:1) and NaBH ₃ CN (1.1 eq) was added. The mixture was stirred until the reaction was complete, quenched by the addition of 20% aqueous sodium hydroxide and extracted several times with diethyl ether. The combined organic phases are dried over Na ₂ SO ₄ , the solvent is removed in vacuo and the crude product is purified by column chromatography on silica (CH ₂ Cl ₂ /MeOH) to give the desired product (41 %) was obtained.
¹ H NMR: (400 MHz, CDCl ₃ ): δ(ppm) = 8.06 (d, 1H, 10.2 Hz), 7.99 (d, 1H, J = 9.9 Hz), 7.24-7.29 (m, 2H), 7.16- 7.21 (m, 1H), 7.11-7.15 (m, 2H) 2.97 (d, 1H, J = 12.3 Hz), 2.77 (s, 3H), 2.63 (s, 3H), 2.54 (d, 1H, J = 6.8 Hz), 2.37-2.47 (m, 5H), 2.26 (s, 3H), 1.95 (t, 2H, J = 11.3 Hz), 1.62-1.78 (m, 4H), 1.48-1.61 (m, 1H), 1.34 -1.44 (m, 2H)
LC/MS: m/z = 444.0, _Rt = 1.16 min (method 4).

3) Biological and pharmacological studies 3.1) Recombinant protein purification 3.1.1) Purification of Strep-tagged TRAF6 RZ1 protein ) and the Ubc13 binding mutant (TRAF6 D57K) were cloned into the pASK-IBA3+ vector (IBA Life Sciences). These proteins were expressed in BL21 codon+RIPL cells (Agilent Technologies) in Luria/Miller medium supplemented with 100 μM ZnCl ₂ , using an AKTA purifier (GE Healthcare Life Sciences) and the Strep- Purification was performed using the Tag®-Strep-Tactin® system.

3.1.2) Purification of untagged TRAF6 RZ1 protein The wild-type TRAF6 RZ1 domain cDNA sequence was cloned into the pGex 4T1 vector (GE Healthcare Life Sciences). For purification of untagged TRAF6 RZ1, protein was expressed in BL21 codon+RIPL cells in M9 mineral medium supplemented with 100 μM ZnCl ₂ . For ¹⁵ N-labeled proteins, normal NH ₄ was replaced with ¹⁵ NH ₄ . Protein purification was performed using Glutathione Sepharose 4 fast flow beads (GE Healthcare Life Sciences), Thrombin clearance (Sigma-Aldrich) and gel filtration chromatography (Superdex 75, GE Healthcare Life Sciences).

3.1.3) Purification of Flag-His-tagged Ubc13 protein (Ubc13-FH) The Ubc13 full-length and Flag-His-tag cDNA sequences were cloned into the pGex 4T1 vector. GST-tagged proteins were expressed in BL21 codon+RIPL cells in LB medium and purified by Glutathione Sepharose 4 fast flow beads, Thrombin clearance and gel filtration chromatography (Superdex 75, GE Healthcare Life Sciences).

3.1.4) Purification of GST proteins (including GST-OTUB1 and GST-Ubc13) was performed according to Weber, E., Rothenaigner, I., Brandner, S., Hadian, K. & Schorpp, K. A High- Throughput Screening Strategy for Development of RNF8-Ubc13 Protein-Protein Interaction Inhibitors. SLAS Discov 22, 316-323 (2017).

3.1.5) His-tagged Uev1a protein was purified according to the GST-OTUB1 protocol with the following buffers: lysis buffer (20 mM NaH ₂ PO ₄ , 20 mM NaCl, 50 mM imidazole), elution buffer (20 mM NaH ₂ PO4, 20 mM NaCl, 500 mM imidazole) and desalting buffer ₍ 20 mM _NaH2PO4 , ₂₀ mM NaCl).

3.2) Identification and Characterization of TRAF6-Ubc13PPI Inhibitors in Biochemical Assays 3.2.1) ALPHAScreen
To identify small molecules that specifically target the RZ1 domain of TRAF6 and thereby prevent binding to Ubc13, ALPHAScreen technology (PerkinElmer) was selected, in high-throughput screening and structure-activity relationship (SAR) analysis, TRAF6 _WT StrepII-Ubc13FH interaction was detected.

To perform the automated ALPHAScreen assay, all of the following components were pre-diluted in ALPHAScreen buffer (1×PBS, 0.5% BSA, 0.01% Tween-20). First, TRAF6 protein (100 nM final concentration, 30 μL volume) was added to a 384-well-Opti-plate containing the MultiFlo distribution system (BioTek), followed by compound transfer through a Sciclone G3 transfer station (PerkinElmer). . Ubc13FH (final concentration 75 nM, 10 μL volume) was added using the MultiFlo system and incubated for 1 hour at room temperature before both beads (Strep-Tactin Alpha donor beads and nickel chelate acceptor beads; final concentration 4 μg each, 10 μL each). volumes) were mixed and added under reduced light using the MultiFlo system. Plate readings were taken after an additional hour of incubation at room temperature. Statistical parameters including coefficient of variation, Z′ value and signal window were calculated to determine the quality of HTS campaigns. To assess compound efficacy, ALPHAScreen units of compound-treated samples were referenced to control DMSO-treated samples. The TRAF6 _D57K mutant thereby served as a minimal signal control and was included on all plates for screening. Compounds that inhibited TRAF6 _WT StrepII-Ubc13FH by more than 25% were considered active substances. After exclusion of ALPHAScreen frequent hitter compounds and His-tag frequent hitters (Schorpp, K., I. Rothenaigner, E. Salmina, et al., Identification of Small-Molecule Frequent Hitters from AlphaScreen High-Throughput Screens. J Biomol Screen, 2014. 19(5): p. 715-26), 178 compounds were defined as primary hits and subsequently tested in a 5-point serial dilution assay (2.5-40 μM) in the TRAF6 _WT StrepII-Ubc13FH ALPHAScreen experiment. did. Only compounds with a dose-dependent effect on TRAF6 _WT StrepII-Ubc13FH (n=27) were considered further. For analog studies, all compounds were analyzed in 10-point increments (0.2-100 μM) in the TRAF6 _WT StrepII-Ubc13FH ALPHAScreen experiment. In parallel, compounds were tested in the ALPHAScreen TruHits Kit (PerkinElmer) to rule out non-specific interference with the ALPHAScreen technology.

さらなるアルファスクリーンの結果を図２０に列挙する。 Table 2: _IC50 values for inhibition of TRAF6-UBC13 interaction.

Further alphascreen results are listed in FIG.

3.2.2) Two-dimensional nuclear magnetic resonance (2D-NMR)
To determine whether inhibitory compounds bind directly to TRAF6 and not to Ubc13, 2D-NMR experiments were performed. Untagged ¹⁵ N-labeled TRAF6 RZ1 protein (c=120 μM) and compound (c=20 mM) were incubated for 10 minutes at a ratio of 1:5 in a 3 mm NMR tube before spectra were acquired. Two-dimensional SOFAST-HMQC spectra were obtained to observe chemical shift perturbations upon compound addition. All measurements were made using a Bruker Avance 600 MHz spectrometer.

3.2.3) In vitro ubiquitination studies 3.2.3.1) Western blot-based assays The ability of TRAF6 to form K63-linked polyubiquitin chains with Ubc13 was analyzed after compound treatment. Therefore, untagged TRAF6 _WT and TRAF6 _D57K were each purified by recombinant techniques. 0.125 μM TRAF6 protein in K63 assay buffer (25 nM Tris-HCl, pH 7.6; 250 nM MgCl ₂ ; 500 nM creatine phosphate; 0.3 U/mL inorganic pyrophosphatase; 0.3 U/mL creatine phosphokinase). , in a total volume of 100 μL, were preincubated with DMSO or compounds for 30 minutes at room temperature. A small aliquot was taken for the input sample. Master containing 0.01 μM E1 activating enzyme (UBE1) (Boston Biochem), 0.2 μM E2 binding enzyme complex (Ubc13/Uev1A) (Boston Biochem), 1 mM ZnCl ₂ , 2 mM ATP and 4 μM monoubiquitin (Boston Biochem) The mixture was added to the TRAF6 protein and the reaction mixture was incubated at 37°C for 120 minutes. Reactions were stopped by adding 4×SDS loading buffer and boiling at 95° C. for 5 minutes. Input samples were analyzed using the Pierce Silver Stain Kit (Thermo Fisher Scientific). Ubiquitination was detected in Western blot analysis using a ubiquitin antibody.

For analysis of cIAP1, MDM2, TRIM63, RNF4, ITCH and E6AP-mediated polyubiquitination, the respective ubiquitin ligase kits from Boston Biochem (K-102, K-260, K-200B, K-220, K- 240 and K-270) were used. Here reaction buffer, E1, E2 and E3 enzymes (and p53 substrate in case of MDM2) are mixed and pre-incubated with compounds or DMSO for 5 minutes at room temperature, followed by Mg ²⁺ -ATP and Mg 2+ -ATP for reaction initiation. Monoubiquitin was added. Ubiquitination was visualized by Western blot analysis.

3.2.3.2) Ubiquitin Profiler (Eurofins Pharma Discovery Services UK)
C25-140 was sent to Eurofins Pharma Discovery Services UK for single point ubiquitin profiler studies of the effects of C25-140 at concentrations of 30 μM against 24 different ubiquitin reaction cascades using distinct combinations of E2, E3 and substrates. gone. Each response was measured in duplicate. Cascade activity is expressed as a percentage of the mean cascade activity in positive control samples. The positive control value is considered 100% and all test samples are referenced to this value.

3.3) Validation of TRAF6-Ubc13 PPI inhibitors in cell-based assays 3.3.1) Electrophoretic-Mobility-Shift-Assay (EMSA) to measure NF-κB activation
For compound treatment, MEF cells were seeded in 6-well plates at 1.2×10 ⁵ cells/well the day before treatment. Jurkat T-cells were seeded immediately prior to treatment at 2×10 ⁵ cells/sample. Six hours after compound or DMSO treatment, cells were stimulated as indicated. MEF cells were thereby left untreated or stimulated with recombinant murine IL-1β (1 ng/mL) or TNFα (10 ng/mL) for 20 minutes. Jurkat T-cells were stimulated with phorbol-12-myristate-13-acetate (PMA) (400 ng/mL) and ionomycin (600 ng/mL) for 30 minutes. Finally, cells were lysed in high salt lysis buffer (20 mM HEPES pH 7.9; 350 mM NaCl; ₁ mM MgCl2; 0.5 mM EDTA; 0.1 mM EGTA; 20% glycerol; 1% NP-40; 1 mM DTT; 300 μM sodium vanadate; complete protease inhibitor) and analyzed by EMSA assay using H2K (NF-κB) or Oct-1 (loading control) probes. The following EMSA oligonucleotides were applied:

3.2. IKK Kinase Assay For MEF cells, 5.5×10 ⁵ cells were seeded, then treated with compounds for 6 hours and finally stimulated with IL-1β or TNFα for 8 minutes. 1×10 ⁶ Jurkat T-cells were treated with compounds for 6 hours and finally stimulated with PMA (400 ng/mL) and ionomycin (600 ng/mL) for 8 minutes. Co-IP lysis buffer (150 mM NaCl; 25 mM Hepes (pH 7.5); 0.2% NP-40; 1 mM glycerol; 10 mM NaF; 8 mM β-glycerophosphate; 1 mM DTT; 300 μM sodium vanadate; ), the input sample was harvested and the supernatant was incubated with 6 μL of the IKKγ antibody FL-419 (Santa Cruz Biotechnology). After incubation with protein-G-sepharose beads (GE Healthcare), the beads were Wash in Co-IP wash buffer (150 mM NaCl; 25 mM Hepes (pH 7.5); 0.2% NP-40; 1 mM glycerol); ^32P -γ-ATP-supplemented kinase reaction buffer (20 μM HEPES pH 7.9; 10 mM MgCl ₂ ; 20 μM ATP; 20 mM β-glycerophosphate; 200 μM sodium vanadate; 1 mM DTT) at 37°C. After 30 minutes of incubation, reactions were stopped and analyzed by autoradiography as well as Western blot after separation on SDS-PAGE.

3.3.3) Analysis of mRNA Expression For MEF cells, 1.2×10 ⁵ cells were seeded per sample, treated with compounds for 6 hours, and stimulated with either IL-1β or TNFα for 60 minutes. Jurkat T-cells and primary mouse CD4 ⁺ T-cells were seeded at 2×10 ⁵ cells per sample, similarly treated with compounds for 6 hours and stimulated with PMA/ionomycin for 3 hours. RNA was isolated according to the manufacturer's protocol of the Qiagen RNeasy Kit (MEF cells) or the STRATEC INVITRAP® SPIN UNIVERSAL RNA MINI KIT (Jurkat T-cells). Isolation of mRNA from epidermal white adipose tissue from DIO mice was performed using TRIzol® reagent according to the manufacturer's instructions. To clean the isolated RNA from genomic DNA, samples (up to 5 μg RNA) were treated with RQ1 RNase-Free DNase I (Promega) as described in the technical manual. For reverse transcription of RNA to cDNA, DNase I-digested RNA samples were processed using the SuperScript III First Strand cDNA Synthesis System (Invitrogen/ThermoFisher Scientific) according to the manufacturer's protocol. Thus, random hexamers provided with the kit were used for reverse transcription. cDNA samples were stored at -20°C. A LightCylcer480 (Roche) and a KAPA SYBR FAST qPCR Kit were used for quantification of cDNA in the samples. For relative quantification, the ΔΔCp method originally described by Pfaffl (Pfaffl, M.W., A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res, 2001. 29(9) : p.e45) was applied. The following primers were used for qRTPCR:

3.3.4) ALPHA Surefire
To detect protein levels of p-IκBα (Ser32/36) as well as total IκBα, 1×10 ⁴ MEF cells were seeded in 96-well plates, treated with compounds for 6 hours, and treated with 1 ng/mL IL-1β. Stimulated for 7 minutes. Protein levels of p-IκBα and total IκBα were analyzed using the ALPHASurefire kit (PerkinElmer) according to the manufacturer's instructions. A general two-plate protocol was performed.

3.3.5) Immunoprecipitation of TRAF6 for detection of endogenous ubiquitination 2.5×10 ⁶ MEF cells were seeded, treated with compounds for 6 hours, and then treated with IL-1β (3.5 ng/mL) for 10 minutes. Stimulated for minutes. Cells were lysed in Co-IP lysis buffer containing 1% SDS to eliminate all cell interactions. After cell lysis and removal of input samples, samples were diluted in 0.1% SDS, incubated with 5 μL of TRAF6 antibody EP591Y (Abcam) and immunoprecipitated with Protein-G-Sepharose beads. After washing the beads in Co-IP wash buffer, TRAF6 protein was eluted and the level of TRAF6 ubiquitination was detected by Western blot analysis.

3.3.6) Enzyme-linked immunosorbent assay (ELISA)
To measure cytokine secretion, 2×10 ⁵ Jurkat T-cells, or primary CD4 ⁺ T-cells, or human PBMCs were seeded per sample, treated with compounds for 6 h, and stimulated as indicated. After 20 hours of stimulation, supernatants were harvested and IL-2, IL-1β and TNFα Ready-Set-Go! provided by Affymatrix eBioscience. Analyzed using an ELISA (2nd Generation) kit.

3.3.7) Co-immunoprecipitation studies in HEK293T-cells To study the interaction of TRAF6 and Ubc13 in cells, 1×10 ⁶ 293T-cells were seeded and 18 hours later subjected to X-tremeGENE HP DNA transfection. Cells were transfected with 1 μg of pEF-HA-empty or pEF-HA-TRAF6 RZ1 plasmid using transfection reagent (Roche). Six hours after transfection, cells were treated with compound or DMSO. After an additional 42 h incubation, cells were harvested in Co-IP lysis buffer and HA-TRAF6 was immunoprecipitated with anti-HA affinity matrix (Roche). Binding of Ubc13 to TRAF6 was examined by Western blot analysis.

3.4) Viability, cell cycle, ADME and PK studies

3.4.1) Viability Assay For cell viability experiments using the CellTiter-Glo® 2.0 system (Promega), MEF cells (1.2×10 ³ cells/well), HepG2 cells (4×10 ³ cells/well) and Jurkat T-cells (5×10 ³ cells/well) were seeded in 384-well plates. After 18 hours, cells were treated with compounds using the Sciclone G3 automated system. After 6 or 24 hours of incubation, luciferase measurements were performed according to the manufacturer's manual.

3.4.2) Cell Cycle Assessment For cell cycle staining, cells were seeded at 2×10 ⁵ cells per sample and treated with compounds or DMSO. After the indicated incubation times (24 or 72 h), cells were fixed in 100% ethanol followed by propidium iodide staining solution (0.1% Triton X-100; 20 μg/ml) for 40 min in the dark. mL propidium iodide; stained with 200 μg/mL RNAse A) in PBS. Propidium iodide fluorescence was collected using an Attune Flow Cytometer (Thermo Fisher Scientific). Analysis was performed using the single cell analysis software FlowJo.

3.4.3) Cell Proliferation Measurement of Osteosarcoma Cell Line U2OS For analysis of cell proliferation of the osteosarcoma cell line U2OS, 5×10 ³ cells were seeded in 96-well plates. After 18 hours, cells were treated with C25-140 (compound 3) or DMSO for 6 hours. A ¹³⁷ cesium source (HWM-D2000) was used to irradiate selected samples with doses of either 2 Gy or 4 Gy. Cell proliferation was measured every 24 hours for 7 days by counting Hoechst 33342 positive cells in an Operetta High-Content Imaging System (PerkinElmer) and Harmony/Columbus analysis software (PerkinElmer).

3.4.4) Determination of cell viability in DLBCL -MYD88wt) and BJAB (GCB DLBCL) cell lines were selected. 1×10 ⁶ cells of each cell line were seeded in 5 mL and treated with 30 μM compound C25-140 (compound 3) or DMSO. The entire experiment was conducted for 9 days. 500 μL were taken daily and viable cell counts were obtained using a ViCEII cell counter (Beckmann Coulter). On days 4 and 8 of the experiment, the volume was adjusted again to 5 mL and samples were supplemented with 30 μM compound C25-140 or DMSO. This process was necessary to ensure linear growth of DMSO-treated cells. All curves for C25-140 treated samples were referenced to corresponding DMSO treated controls.

3.4.5) ADME and Pharmacokinetic Studies All ADME (plasma stability, plasma protein binding, microsomal stability, LogD, Caco-2 assay, CYP450 inhibition and hERG Predictor assays) of C25-140 (compound 3), and pharmacokinetic studies (T _max , C _max , AUC _{0-240 min} , mean residence time, elimination half-life, elimination rate constant, volume of distribution and clearance) were performed by Bienta/Enamine Ltd. Bienta Enamine Biology Services, Inc.

3.4.6) Safety Screen 87
A total of 87 assays, SafetyScreen 87, were performed at Eurofins Cerep Panlabs. 87 primary molecular targets, including 13 enzymatic assays (kinase and non-kinase enzymes) and 74 binding assays (nuclear receptors, GPCRs, transporters and ion channels), were performed at a compound concentration of 10 μM C25-140. . Each of the 87 assay reactions was performed with optimized conditions historically available at Eurofins Cerep Panlabs. Reference compounds for each of the 87 reactions were tested concurrently in parallel with C25-140 to ensure optimal assay setup. In this study, the reference compounds performed in the same range as the historical Ki values.

3.4.7) Omniscreen
The effect of C25-140 on cell viability of 80 cancer cell lines using the CellTiter-Glo Luminescent Cell Viability Assay was investigated by CrownBioscience Inc. conducted at our Beijing test facility. Here, 80 selected cell lines were cultured at 37° C., 5% CO ₂ and 95% humidity in medium supplemented with 10-15% FBS. Culture media were purchased from GIBCO or Sigma (USA). Cells were treated with C25-140 at 9 dose levels (3.16-fold serial dilution starting at 80 μM). Cisplatin served as a reference control and was started at the highest working concentration of 100 μM. Test samples were incubated for 72 hours and cell viability was measured by CellTiter-Glo assay.

3.5) Mouse studies

3.5.1) Isolation of primary mouse CD4-positive T-cells CD4 ⁺ T-cells from peripheral lymph nodes of Balb/C mice were isolated using the CD4 ⁺ T-cell isolation kit II (MILtenyi). was performed by negative magnetic activated cell selection (MACS). Primary CD4 ⁺ T-cells were treated with 10% heat-inactivated FCS, 1% penicillin/streptomycin, 1% NEAA (Life Technologies), 1% HEPES, 1% L-glutamine, 1% sodium pyruvate (Life Technologies) and 0. .Cultivated in RPMI medium supplemented with 1% β-mercaptoethanol. 2×10 ⁵ cells were seeded per sample and treated with compounds for 6 hours. Cells were stimulated with anti-CD3 (0.5 mg/mL) and anti-CD28 (1 mg/mL) on plates pre-coated with rabbit anti-hamster IgG or anti-CD3/CD28 coated beads (3:1). Samples for qRT-PCR were collected after 4 hours, while supernatants for analysis of cytokine secretion were collected after 20 hours of stimulation.

3.5.2) Diet-Induced Obesity (DIO) Mouse Study Mice were fed a high-fat diet until their body weight reached at least 40 g to develop diabetes-induced obesity. These mice were divided into a control group and a group for C25-140 (compound 3) injection. Mice were treated daily with 14 μmol/Kg (=7.4 mg/Kg) of C25-140 intraperitoneally for 20 days. To this end, C25-140 was dissolved in 2.9% DMSO, 1.5% Tween-80 and 1x PBS. Body weight and food intake were measured daily prior to injection. On day 20, mice were sacrificed 2 hours after compound injection and epidermal white adipose tissue was removed from both body sites, frozen in liquid nitrogen and stored at -80°C.

3.5.3) Psoriasis Mouse Studies To evaluate the anti-inflammatory activity of Compound C25-140 (Compound 3), an imiquimod (IMQ)-induced psoriasis mouse model was performed by Washington Biotechnology, Inc. (Washington Biotechnology, Inc.). (Baltimore, USA). Per group, 8 male Balb/C mice were shaved on the back (1.5 cm x 2 cm) and IMQ and C25-140 were applied topically to the shaved back and right ear daily for 6 days. For that, C25-140 was dissolved in acetone and applied at a total dose of 500 μg/day (2×250 μg). Mice were monitored and daily scores (erythema, scaling and dorsal skin thickness) were collected independently on a scale of 0-4: 0=none; 1=slight; 2=moderate; 4 = very noticeable. Ear thickness was measured by an electronic caliper as an index of edema. IL-17 cytokine secretion in right ear tissue was measured by ELISA.

3.5.6) Collagen-Induced Arthritis Model To test the potential of C25-140 to ameliorate RA symptoms, a collagen-induced arthritis (CIA) model in DBA1/J mice was developed by Washington Biotechnology, Inc. (Washington Biotechnology, Inc.). (Baltimore, USA). Each group of 10 mice received a single subcutaneous injection of collagen/complete Freund's adjuvant emulsion (0.05 mL/mouse; 100 μg collagen/CFA). Twenty days later (day 21), mice were boosted with collagen (0.05 mL/mouse; collagen in incomplete Freund's adjuvant, 100 μg/mouse). On day 28, mice were scored for their arthritis index (AI) and were intraperitoneally administered C25-140 at 6 mg/Kg, 10 mg/Kg and 14 mg/Kg, respectively, twice daily for 14 days. . Mice were scored daily for gross signs of arthritis. To that end, the following scoring indices were applied to individual paws: 0 = no apparent effect of arthritis; 1 = single digit edema and/or erythema; 2 = double digit edema and/or erythema; 3 = 2. Greater than one order of magnitude of edema and/or erythema; 4 = severe arthritis throughout feet and toes. A single forelimb score was added and recorded. On day 42, animals were euthanized, limbs harvested and histopathology performed using toluidine blue staining along with scoring of several parameters.

3.6) Human Donor Experiments To isolate peripheral blood mononuclear cells (PBMC) from 50 mL of human blood from 3 different donors, the blood was mixed with 800 units of heparin (Sigma-Aldrich) and subjected to 300 xg for 10 minutes. Centrifuge without brake. After removal of the plasma fraction, the buffy coat was diluted with 2 volumes of PBS and added on top of 15 mL of Lymphoprep (STEMCELL technologies). After centrifugation at 160xg for 20 minutes at room temperature without brake, platelets were removed and the cell suspension was centrifuged again at 350xg for 20 minutes without brake. Mononuclear cells from the middle layer were transferred to new falcons and washed three times with PBS supplemented with 0.1% BSA and 2 mM EDTA. Finally, cells were resuspended in RPMI medium containing 10% FCS, 1% penicillin/streptomycin and 50 μM β-mercaptoethanol. For each sample, 2×10 ⁵ cells were seeded and treated with compound for 6 hours. Cells were treated with 1 μg/mL lipopolysaccharide (LPS), or CD3/CD28 (1 μg hCD3 (IgG _2A ), 4 μg hCD28 (IgG ₁ ), 2 μg ant-IgG _2A, and 2 μg ant-IgG _2A ), or 20 ng/IL-1β. Supernatants for analysis of cytokine secretion were harvested 20 hours after stimulation.

3.7) Statistical Analysis Experiments are presented as mean±standard deviation of the mean. For statistical analysis, the 'unpaired t-test' was applied and statistical significance was determined by p-value < 0.05. In the DIO-mouse study, statistical significance of weight loss was calculated with a two-way Anova test. A two-way Anova test was applied to determine whether C25-140 (compound 3) treatment significantly affected the outcome of psoriasis symptoms. Finally, a two-way Anova test was applied to DLBCL experiments focused on survival curves. Statistical parameters were determined using Prism 6.0 software (GraphPad)

4) Discussion In a high-throughput screening (HTS) campaign to identify small molecules that inhibit the TRAF6-Ubc13 interaction, C25 (equivalent to compound 1 herein) is in the process of using in vitro and cell-based assays. It was found as a promising hit in a systematic process (Fig. 1A). A structure-activity relationship (SAR) study centered on C25 revealed three compounds with similar in vitro potency (FIGS. 1B and 2A), whereas C25-140 (equivalent to compound 3 herein) It contained improved inhibitory activity in a cell-based NF-κB activation assay (Fig. 1C). Moreover, C25-140 dose-dependently decreased the interaction between endogenous Ubc13 and ectopically expressed TRAF6 in cells (FIG. 2B). Further characterization of C25-140 in vitro experiments showed that this compound directly binds to the TRAF6 protein (Fig. 2C), as evidenced by NMR studies, and C25-140 is the TRAF6 E3 ligase that forms polyubiquitin chains. We showed that activity can be decreased in a dose-dependent manner (Fig. 2D). Importantly, C25-140 did not affect Ubc13 binding to Uev1a (FIG. 2E). Thus, C25-140 has the ability to bind TRAF6 and subsequently interfere with its binding to Ubc13, ultimately reducing TRAF6 activity.

Before extending detailed cellular studies of C25-140, we first analyzed whether this compound affects cell viability and cell cycle phases. Here, C25-140 did not significantly affect cell viability in three different cell lines (Jurkat, MEF and HepG2) except at the highest concentration of 100 μM (Fig. 3A). Furthermore, no effect on cell cycle phases was observed with C25-140 treatment of Jurkat and U2OS cells (FIGS. 3B-C). Overall, we demonstrate that C25-140 did not affect cell viability and cell cycle progression at concentrations up to 30 μM used for subsequent cell-based experiments. can be done.

As an initial cell model, we chose MEF cells that can be stimulated with IL-1β and TNFα. Stimulation of these cells with IL-1β induces TRAF6 autoubiquitination, which is required for signal progression towards IκBα phosphorylation by the IKK complex, activation of NF-κB, and expression of NF-κB-dependent target genes. . C25-140 treatment of MEF cells resulted in decreased TRAF6 autoubiquitination (Fig. 4A), reflecting the consequences of in vitro ubiquitination by the recombinant protein (Fig. 2D). Decreased TRAF6 autoubiquitination was followed by decreased IKK kinase activity (Fig. 4B), which led to decreased target gene expression (Fig. 4C). These data show that inhibition of TRAF6 activity is seen early in IL-1 receptor signaling across various stages of signaling and that inhibition of TRAF6 activity can be visualized at all these different stages. . Surprisingly, TNFα-induced phosphorylation of IκBα (FIG. 4D) and NF-κB-induced target gene expression (FIG. 4E) were also affected by C25-140 treatment. This was unexpected since, to the best of our current knowledge, TRAF6 is largely implicated in signal progression in TNFα receptor signaling. To clarify this finding, we tested the effect of C25-140 on the activity of several other E3 ligases to understand if there is any cross-reactivity. These studies revealed that the E3 ligase activity of cIAP1, an E3 ligase that generates K63-linked polyubiquitin chains similar to TRAF6, is indeed influenced by C25-140 (Fig. 5A). In contrast, several other E3 ligases (MDM2, TRIM63, ITCH, E6AP and RNF4) that assemble different types of polyubiquitin chains (mainly K11 and K48) were unaffected by C25-140 ( 5B-D). In addition, 24 additional ubiquitination reactions involving different E3-E2 complexes and substrate combinations were tested in the Ubiquitin Profiler study. None of these responses were inhibited by C25-140 (Fig. 5E). Since the E3 ligase cIAP1 is a central regulator of TNFα signaling that is inhibited by C25-140, these data explain the inhibitory effect of C25-140 on TNFα signaling. Very importantly, these data also show that C25-140 is not a broadband E3 ligase inhibitor and thus retains good selectivity (although minor off-target effects on TNFα signaling are observed). ).

We also tested C25-140 on T-cell receptor (TCR) signaling in a human T-cell line (Jurkat T-cells) of the adaptive immune response. Here we can show that C25-140 also affects TRAF6-dependent TCR signaling (Oeckinghaus, A., E. Wegener, V. Welteke, et al., Malt1 ubiquitination triggers NF-kappaB signaling upon T-cell activation. EMBO J, 2007. 26(22): p. 4634-45). In Jurkat T-cells, TCR stimulation resulted in TRAF6 autoubiquitination, which was dose-dependently decreased by C25-140 (Fig. 6A). Decreased TRAF6 autoubiquitination resulted in decreased downstream events including IKK kinase activity, NF-κB activation and expression and secretion of the respective cytokines (FIGS. 6B-E). These data emphasize that our compounds also affect TCR-mediated adaptive immune responses in human-derived cell lines.

After showing that C25-140 successfully competes with TRAF6-dependent receptor signaling in mouse and human cell lines, we next turned to more biologically relevant cells. For this purpose, we isolated human peripheral blood mononuclear cells (PBMC) from three different individuals and stimulated these cells with LPS to activate the innate immune response. Consistent with our previous data, C25-140 regulates the secretion of NF-κB-driven inflammatory cytokines (TNFα and IL-6 after IL-1β stimulation (Fig. 7A), IL-1β and TNFα (Fig. 7B)), indicating that C25-140 not only affects TRAF6 activity in immortalized cells, but more importantly, innate immune processes in primary human blood cells. decreases. A similar inhibitory effect by C25-140 could be observed in human PBMC by stimulating the adaptive immune response with CD3/CD28 (Fig. 7C). Finally, we also isolated primary mouse CD4 ⁺ T-cells for progenitor experiments to in vivo mouse studies. C25-140 also prevented CD3/CD28-mediated NF-κB activation and IL-2 production (FIG. 8).

Taken together, data from in vitro and cell-based studies involving immortalized cell lines and primary human and mouse cells demonstrate that C25-140 exerts inhibitory effects on a variety of immune and inflammatory signaling pathways. and show that this may be beneficial for treating a highly inflamed immune system.

To move on to in vivo mouse studies, we analyzed whether compound C25-140 has suitable properties for in vivo applications. To this end, we first performed ADME studies examining the following parameters: (i) plasma stability, (ii) plasma protein binding, (iii) microsomal stability, (iv) logD , (v) Caco-2 permeability, (vi) CYP450 inhibition and (vii) hERG inhibition. Compound C25-140 contained a fairly good set of properties in all of these assays (Figure 9A). Overall, the compound is stable in plasma and microsomes, exhibits good penetration of the Caco-2 layer indicating good oral availability, has adequate and good plasma protein binding capacity, and very important In particular, it does not significantly inhibit hERG. Only CP450 has some inhibitory potential against selected CYPs.

We also tested C25-140 pharmacokinetics (PK) after intravenous (IV), oral (PO) and intraperitoneal (IP) injection of 10 mg/Kg of each compound. All analytical data are summarized in Figure 9B and a concentration/time course graph of IP administration is shown in Figure 9C. When administered IP, C25-140 showed a higher calculated in vivo half-life in the circulation (184 min) compared to IV (80 min) and PO (127 min) delivery. Overall, a rapid initial distribution phase is observed. This compound has a good calculated oral bioavailability of about 45%. No apparent adverse effects of the compound on animals were observed during the PK study.

Subsequently, we applied the Cerep SafetyScreen 87 panel (a suite of key receptors, proteins and enzymes that should not be affected to ensure good in vivo tolerance) to identify C25- 140 were tested for safety. Interestingly, no significant binding of C25-140 was detected to any of these targets (Figure 10), suggesting that C25-140 has a high degree of safety for preclinical/clinical applications. clearly suggesting that

In initial mouse studies, we wanted to explore the potential of the TRAF6 activity inhibitor C25-140 to prevent inflammation and weight gain caused by high-fat diet (HFD)-induced obesity. In collaboration with the Diabetes and Obesity Institute of the Helmholtz Zentrum München GmbH-Deutsches Forschungszentrum fur Gesundheitun Umwelt (HMGU), 16 mice were selected which were fed HFD for at least 12 months. These mice had an average body weight of 55 g and had obvious features of obesity and type 2 diabetes (T2D). These mice were divided into two groups and treated intraperitoneally once daily with either vehicle or C25-140 at a dose of 7.4 mg/Kg for 20 days. Mice were monitored for body weight and their food intake daily prior to injection. Control-treated mice continuously gained weight under HFD over 20 test days, whereas C25-140-treated mice gained little weight (FIG. 11A). Importantly, food intake did not differ between the two groups (Fig. 11B), indicating that treatment had no adverse effects on mice in general. We also examined isolated adipocytes for IL-1β expression as a marker of inflammation and VCAM as an NF-κB target gene. Both genes were significantly decreased with treatment, providing molecular evidence that the TRAF6 inhibitor indeed functioned correctly. Overall, this study provides important information that the TRAF6 inhibitor C25-140 can reduce inflammation in vivo, thereby contributing significantly to reduced weight gain under high-fat diet conditions. .

A second in vivo mouse model was conducted by CRO Washington Biotechnology aimed at investigating C25-140 in psoriasis mouse studies as an autoimmune disease model. In this model, psoriasis is triggered by imiquimod (IMQ) via activation of TLR7 signaling (Gilliet, M., C. Conrad, M. Geiges, et al., Psoriasis triggered by toll-like receptor 7 agonist imiquimod in the presence of dermal plasmacytoid dendritic cell precursors. Arch Dermatol, 2004. 140(12): p. 1490-5). More specifically, IMQ was applied once daily to the shaved back and right ear of mice. C25-140 was topically applied twice daily to the same site. Parameters capturing disease outcome were scored daily and samples for IL-17 cytokine measurement were taken on day 6 (FIG. 12A). Compound 25-140 was able to significantly reduce the 'cumulative score' (Figure 12B) and 'thickness score' (Figure 12C). These scores reflected individual scores such as "desquamation" (Fig. 12D), "erythema" (Fig. 12E) and "ear thickness" (Fig. 12F). All of these parameters reflected improvement in disease symptoms. Finally, analysis of right ear tissue for IL-17 revealed that this cytokine was significantly reduced in its secretion/expression (Fig. 12G). Thus, this study demonstrated that C25-140 as an inhibitor of TRAF6 activity can ameliorate symptoms in an in vivo disease model of autoimmune psoriasis.

In a third in vivo mouse study, we tested whether compound C25-140 was effective in a preclinical mouse model of autoimmune rheumatoid arthritis (RA). Here, we tested C25-140 in the collagen-induced arthritis (CIA) mouse model, the most commonly used model of RA efficacy (Brand, D.D., K.A. Latham, and E.F. Collagen-induced arthritis. Nat Protoc, 2007. 2(5): p. 1269-75). This study was also performed by CRO Washington Biotechnology. Here, rheumatoid arthritis (RA) was induced on day 0 by injection of collagen. Twenty-one days later, mice received a collagen booster injection to induce the development of arthritic symptoms. On day 28, C25-140 was administered twice daily at three different doses (6 mg/Kg, 10 mg/Kg and 14 mg/Kg) for 14 days. Prednisolone served as a positive control. Mice were scored daily for arthritis index (AI) and euthanized on day 42. Limbs were harvested for histopathological analysis (Fig. 13A). Body weight was monitored throughout the study and showed no signs of loss (Fig. 13B). Excitingly, our compound almost completely improved the arthritic index to baseline levels at doses of 10 mg/Kg and 14 mg/Kg in this efficacy model (Figure 13C). More importantly, these results were supported by limb histology. Quantification of various parameters (total score, inflammation, pannus, cartilage damage and bone resorption) of disease appearance (Fig. 13D-E) (Fig. 13F) and histological sections of limbs showed that C25-140 dose-dependently reduced RA symptoms. proved to be substantially improved. This entire data set demonstrates that C25-140 successfully resolved the clinical symptoms of RA and reversed the disease state. We present here a successful proof of concept study for compound C25-140 in a preclinical RA efficacy model.

We further investigated the possibility that C25-140 selectively kills a subset of aggressive ABC DLBCLs. ABC DLBCL survival depends on a variety of different mutations that lead to constitutively active NF-κB signaling. The major mutations are within the B-cell receptor (BCR) and MYD88 signaling (Nagel, D., M. Vincendeau, A.C. Eitelhuber, et al., Mechanisms and consequences of constitutive NF-kappaB activation in B-cell Oncogene, 2014. 33(50): p. 5655-65 and Ngo, V.N., R.M. Young, R. Schmitz, et al., Oncogenically active MYD88 mutations in human lymphoma. Nature, 2011. 470(7332): 115-9). To date, most available therapies focus on the BCR pathway. Since TRAF6 is a major player in MYD88 signaling, we wanted to understand if our TRAF6 inhibitors would selectively kill ABC DLBCLs with constitutively active MYD88 mutations. I tried We used MYD88-dependent ABC DLBCL (HBL-1 with MYD88 L265P mutation), MYD88 wild-type ABC DLBCL (U2932) and GCB DLBCL (BJAB) cell lines. All cell lines were treated with C25-140 on days 0, 4 and 8. The study was conducted for 9 days and cell viability (cell number) was measured daily. C25-140 significantly reduced the viability of MYD88-dependent HBL1, whereas MYD88-independent U2932 cells were only slightly affected (FIG. 14). The unrelated GCB DLBCL cell line BJAB did not show any difference in viability with C25-140 treatment (Figure 14). These data indicate that TRAF6 C25-140 is a selective inhibitor of MYD88-dependent ABC DLBCL and may serve as a key inhibitor for combination treatment with current BCR-acting therapeutics.

In the second cancer context, we investigated whether our TRAF6 inhibitor C25-140 was effective against osteosarcoma-derived cells and could be used for osteosarcoma treatment. investigated. We treated U2OS cells with either irradiation (2 Gy or 4 Gy), or C25-140 alone, or a combination of C25-140 and 2 Gy irradiation. C25-140 alone and 2 Gy irradiation had the same inhibitory effect on cell proliferation of U2OS cells, but the combination of both enhanced the effect in the area of 4 Gy irradiation (Fig. 15). These data demonstrate that C25-140 can enhance the efficacy of radiotherapy in such a way that even with less exposure to radiation, its combination with C25-140 ensures effective treatment.

To get a more comprehensive view of the effects of C25-140 on cancer cells, we initiated a cancer study using 78 cancer cell lines of different organ origins. These cancer cell lines were tested against nine concentrations of C25-140. The results indicate that the main effect is on blood-borne cancer cells (Fig. 16). However, cancer cell lines from other tissues were also affected, including bladder, colorectal, liver, lung, pancreas, pharynx and prostate (Figure 16). Importantly, the two control cell lines HS-5 and MCF 10A were completely unaffected, suggesting no general signs of toxicity (FIG. 16).

In the process of identifying compounds that are not commercially available and have unique structures, compounds were synthesized and tested in the ALPHAScreen assay for their ability to interfere with TRAF6-Ubc13 binding. At the same time, these compounds were tested with the TruHits kit to eliminate compounds that interfere with the ALPHAScreen technology. All novel compounds inhibited the TRAF6-Ubc13 interaction, with IC50s varying from 2.3 μM to 19.6 μM (FIG. 17). Neither compound interfered with the TruHits assay. These data demonstrate the existence of novel TRAF6-Ubc13 binding inhibitors with novel structures that are not commercially available. Importantly, all compounds exhibited similar or better toxic behavior compared to C25-140. Up to 50 μM, all compounds showed little toxic effect on three different cell lines (Figure 18). Only at a compound concentration of 100 μM could some toxicity be detected for a distinct compound. Finally, all of these novel compounds were tested for their ability to interfere with TRAF6-Ubc13 binding in a cell-based co-immunoprecipitation assay. In addition to C25-140, further novel compounds HZM06-1 (equal to compound 28), HZM06-2 (equivalent to compound 29), HZM06-3 (equal to compound 30), HZM06-4 (equal to compound 31), HZM06-6 (equal to Compound 32), HZM06-7 (equal to Compound 33), HZM06-8 (equal to Compound 34), HZM06-9 (equal to Compound 35) and HZM06-10 (equal to Compound 36) are also Also active in cells, C25-140, HZM06-4, HZM06-7, HZM06-8 and HZM06-9 were the most active analogs (Figure 19). Taken together, the data in Figures 14-16 demonstrate a series of novel, commercially unavailable compounds with in vitro and cellular activity and little toxicity.

Additional new compounds were synthesized and tested in the ALPHAScreen assay. These compounds showed inhibitory effects to different extents as shown in the table (Figure 20).

It will be appreciated that the invention may be practiced otherwise than as specifically described in the foregoing description and examples. Many modifications and variations of the present invention are possible in light of the above teachings and thus fall within the scope of the appended claims.

Claims (13)

A pharmaceutical composition for use in the treatment of cancer, immune disorders, Parkinson's disease, cardiac hypertrophy or type 2 diabetes, comprising formula I:

(In the formula,
X is selected from the group consisting of N and CH;
R ¹ is selected from the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl, and (C ₆ -C ₁₀ )aryl;
R ² and R ³ are independently selected from the group consisting of (C ₁ -C ₆ )alkyl;
n is an integer between 1 and 2;
Z is C=O;
A is —N(R ⁴ )(R ⁵ ) and the following formula:

selected from the group consisting of
R ⁴ is hydrogen or (C ₁ -C ₆ )alkyl;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, which are optionally halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ₆ ;
R ⁶ is (C ₁ -C ₄ )alkyl;
Y is selected from the group consisting of NB, CH-B and O;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, and (C ₁ -C ₆ )alkyl, which are optionally selected from the group consisting of halogen, (C ₁ -C ₄ )alkyl, and —OR ⁶ , substituted by one or more identical or different substituents;
p is an integer between 1 and 2;
q is an integer between 1 and 3)
or a pharmaceutically acceptable salt, solvate or hydrate thereof. 2. A pharmaceutical composition for use according to claim 1, wherein said compound of formula (I) is represented by formula (II):

(In the formula,
X is selected from the group consisting of N and CH;
^R1 is
i) the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl and (C ₆ -C ₁₀ )aryl, or ii) hydrogen, (C ₁ -C ₆ )alkyl, (C ₃ -C ₈ )cycloalkyl, and (C ₆ -C ₁₀ )aryl;
R ² and R ³ are independently selected from the group consisting of (C ₁ -C ₆ )alkyl;
n is i) an integer between 1 and 2, or
ii) is 2;
Z is selected from the group consisting of C=O;
Y is N or CH;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, and (C ₁ -C ₆ )alkyl,
These are sometimes
i) the group consisting of halogen, (C ₁ -C ₄ )alkyl, and —OR ⁶ ;
or ii) substituted by one or more identical or different substituents selected from the group consisting of halogen and ^OR6 ;
p is an integer between 1 and 2;
q is an integer between 1 and 3)
or a pharmaceutically acceptable salt, solvate or hydrate thereof. 3. A pharmaceutical composition for use according to claim 1 or 2, wherein said compound having a structure according to formula (I) or formula (II) is characterized in that X is N Composition. A pharmaceutical composition for use according to any one of claims 1 to 3, wherein the compound having a structure according to formula (I) or formula (II) is
R ¹ is selected from the group consisting of hydrogen, (C ₁ -C ₄ )alkyl, (C ₃ -C ₆ )cycloalkyl, and (C ₆ -C ₁₀ )aryl;
R ² and R ³ are independently selected from the group consisting of (C ₁ -C ₄ )alkyl and (C ₁ -C ₄ )haloalkyl;
n is
i) an integer between 1 and 2, or ii) 2;
and R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₁₀ )heteroaryl( C ₁ -C ₆ )heteroalkyl, which are optionally
i) one or more identical or different substituents selected from the group consisting of i) halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and —OR ⁶ or ii) the group consisting of halogen A pharmaceutical composition, characterized in that it is replaced by A pharmaceutical composition for use according to any one of claims 1 to 4, wherein said compound of formula (I) is represented by formula (III):

(In the formula,
n is
i) an integer between 1 and 2, or ii) 2;
Y is N or CH;
B is (C ₆ -C ₁₀ )aryl, (C ₃ -C ₁₀ )heteroaryl, (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkyl, which are optionally
one or more identical selected from i) the group consisting of halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )haloalkyl and -OR ⁶ or ii) the group consisting of halogen and -OR ⁶ or substituted by different substituents;
q is
i) an integer between 1 and 3,
ii) an integer between 1 and 2, or iii) is 2)
or a pharmaceutically acceptable salt, solvate or hydrate thereof. A pharmaceutical composition for use according to any one of claims 1 to 5, wherein said compound of formula (I) is represented by formula (IV):

(In the formula,
X is selected from the group consisting of N and CH;
^R1 is
i) the group consisting of hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )haloalkyl, (C ₃ -C ₈ )cycloalkyl and (C ₆ -C ₁₀ )aryl;
ii) selected from the group consisting of hydrogen and (C ₁ -C ₆ )alkyl, or iii) the group consisting of (C ₁ -C ₆ )alkyl;
R ² and R ³ are independently selected from the group consisting of (C ₁ -C ₆ )alkyl;
n is i) an integer from 1 to 2, or ii) 2
is;
Z is C=O;
^R4 is
i) hydrogen or (C ₁ -C ₆ )alkyl, or ii) hydrogen;
R ⁵ is (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )alkyl, (C ₃ -C ₁₀ )heteroaryl(C ₁ -C ₆ )heteroalkyl and (C ₆ -C ₁₀ )aryl(C ₁ -C ₆ )heteroalkyl, optionally i) halogen, (C ₁ -C ₄ )alkyl, ( C ₁ -C ₄ ) substituted by one or more identical or different substituents selected from the group consisting of haloalkyl and —OR ⁶ or ii) halogen;
R ⁶ is (C ₁ -C ₄ )alkyl)
or a pharmaceutically acceptable salt, solvate or hydrate thereof. A pharmaceutical composition for use according to any one of claims 1 to 6, wherein the cancer is a lymphoma such as diffuse large B-cell lymphoma (DLBCL) and MALT lymphoma, multiple myeloma (MM). ), lung cancer, lung adenocarcinoma, colon cancer, prostate cancer, breast cancer, osteosarcoma, pancreatic cancer and esophageal squamous cell carcinoma (ESCC). Pharmaceutical composition for use according to any one of claims 1 to 6, wherein the immune disease is an autoimmune disease 9. The pharmaceutical composition of claim 8, wherein the autoimmune disease is from psoriasis, rheumatoid arthritis, celiac disease, inflammatory bowel disease such as Crohn's disease and ulcerative colitis, multiple sclerosis and diabetes type 1. A pharmaceutical composition selected from the group consisting of

A compound selected from the group consisting of 11. A pharmaceutical composition comprising a compound of claim 10 and at least one pharmaceutically acceptable carrier. 12. A kit comprising the pharmaceutical composition of claim 11 and at least one pharmaceutically acceptable carrier. The compound is

2. The pharmaceutical composition of claim 1, selected from the group consisting of:

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